Brain Research Protocols 1 Ž1997.

203–209

Protocol

Male rat sexual behavior

˚
Anders Agmo )

Laboratoire de Psychophysiologie, Faculte´ des Sciences, UniÕersite´ de Tours, Parc de Grandmont, F-37200 Tours France
Accepted 2 May 1996

Abstract

The male rat’s sexual behavior constitutes a highly ordered sequence of motor acts involving both striate and smooth muscles. It is
spontaneously displayed by most adult male rats in the presence of a sexually receptive female. Although the behavior is important for the
survival of the species it is not necessary for survival of the individual. In that way it is different from other spontaneous behaviors such
as eating, drinking, avoidance of pain, respiration or thermoregulation. Among other things, this means that it is difficult to talk about
sexual deprivation or need. Nevertheless, studies of male sex behavior distinguish sexual motivation Žthe ease by which behavior is
activated, ‘‘libido’’. from the execution of copulatory acts Žperformance, ‘‘potency’’. ŽMeisel, R.L. and Sachs, B.D., The physiology of
male sexual behavior. In: E. Knobil and J.D. Neill ŽEds.., The Physiology of Reproduction, 2nd Edn., Vol. 2, Raven Press, New York,
1994, pp. 3–105 w13x..
The hormonal control of male sexual behavior has been extensively studied. It is clear that steroid hormones, androgens and estrogens,
act within the central nervous system, modifying neuronal excitability. The exact mechanism by which these hormones activate sex
behavior remains largely unknown. However, there exists a considerable amount of knowledge concerning the brain structures important
for sexual motivation and for the execution of sex behavior. The modulatory role of some non-steroid hormones is partly known, as well
as the consequences of manipulations of several neurotransmitter systems.
q 1997 Elsevier Science B.V. All rights reserved.

Themes: Neural basis of behavior

Topics: Motivation and emotion; Hormonal control of reproductive behavior; Psychopharmacological agents

Keywords: Reproduction; Hormone; Psychopharmacology; Neurotransmitter

1. Type of research sensorimotor coordination, elaboration and interpreta-

Ø Studies of male sexual behavior are particularly suitable
for analyses of mechanisms, behavioral and neurobio-
logical, of motivation in the absence of deprivation or
physiological need. This may constitute an important
advantage in relation to many other motivated behav-
iors, like feeding or drinking, where motivation is
activated by deprivation, something that is incompatible
with animal comfort.
Ø Obviously, sex behavior is ideally suited for studies of
the relationship between steroid hormone action at the
cellular level and their behavioral consequences.
Ø Because male sexual behavior is activated by stimuli
emitted by the female it is well suited for studies of
)
Corresponding author. Fax: q 33 47367040;
agmo@balzac.univ-tours.fr
tion of sensory information, etc.
Ø The field of behavioral pharmacology is showing an
increasing interest in male rat sexual behavior as a
model for evaluating drug actions.
Ø Last, but not least, the neurobiology of sexual behavior
is a field of intrinsic importance. Adequate sexual
functioning is an immeasurable source of human well-
being and fundamental for animal production. By ex-
tension, dysfunctional sexual behavior is a frequent
origin of psychological problems in the human and may
adversely affect the reproductive potential of domestic
animals.
2. Time required
E-m ail: Most studies of male rat sexual behavior use sexually
experienced rats. Therefore, the time needed for allowing

1385-299Xr97r$17.00 Copyright q 1997 Elsevier Science B.V. All rights reserved.

PII S 1 3 8 5 - 2 9 9 X Ž 9 6 . 0 0 0 3 6 - 0
204 ˚
A. Agmor Brain Research Protocols 1 (1997) 203–209
the animals to acquire limited but sufficient sexual experi-
ence is included.
Ø A. Pair housing of 20 rats: 15 min.
Ø B. Habituation to a reversed lightrdark cycle: 3 weeks.
Ø C. Three screening tests with receptive female: 7.5 h.
Ø D. Testing 16 animals Žtest ended at the first postejacu-
latory interval.: 2 h.
Ø E. If tests are recorded on video, and a large number of
behavioral items are to be registered, a separate scoring
session is needed: 5 h.
This estimate is limited to the behavioral procedure. If
other manipulations, such as drug injection Žsystemic or
intracerebral., microdialysis, voltammetry, brain lesions,
etc. are to be performed, separate time estimates for these
need to be made.
Since heterosexual behavior requires individuals of dif-
ferent sex, it is also necessary to prepare sexually receptive
females.
Ø A. Ovariectomy of 6 females and implantation of Silas-
tic capsule: 1 h.
Ø B. Preparation of steroid hormone solutions: 1.5 h.
Ø C. Injection of the females with estrogen and 48 h later
with progesterone: 10 min Žthis needs to be repeated
before each test..
Intact females can be used. In that case, step A is
omitted. If ovariectomized females are implanted with
estrogen-containing Silastic capsules, only progesterone
needs to be injected.
3. Materials
Adult Ž3 months or minimum 250 g. male rats are
normally used. Older rats are also vigorous copulators. The
intensity of sexual behavior does not begin to decline until
about 11 months of age w19x. Animals older than 1 year are
rarely used, if the purpose is not to analyze aging and sex
behavior. The strain does not seem to be of great impor-
tance. We normally use Wistar rats, but the Sprague–Daw-
ley and Long–Evans strains are sometimes preferred.
However, there are no systematic studies on strain differ-
ences and sex behavior. Females used as mating partners
are normally of the same strain as the males and should
not be older. The animals are almost always housed under
a reversed lightrdark cycle. Sexual behavior is somewhat
more intense during the dark phase, although sexually
experienced male rats readily copulate close to a sunny
window on a summer afternoon. A 12 h lightr12 h dark
cycle is frequently used, but some prefer 14 h light and 10
h dark. The exact duration of dark Žor light. seems to be
without importance. It is convenient to have the light
turned off at 09.00 or even later so that the animals can be
fed and cleaned before the beginning of the dark phase.
Females are usually kept in the same room as the males.
Food Žany standard rat pellets. and water are always
available. During the dark phase, a dim white light Ž10–25
lux. may constantly be left on to permit handling of the
animals.
We house the animals in pairs, basically for their
comfort. Larger cages with 5 or 10 animalsrcage are more
practical if the animals need to be transported between the
animal quarters and the observation room and require less
space. Animals can also be housed singly. There is no
evidence that housing condition is a determinant of sexual
behavior.
3.1. Special equipment
The test arena may be a rectangular wooden box with
Plexiglas front Ž40 = 60 = 40 cm high. and wire mesh top,
or a Plexiglas cylinder, diameter 60 cm, height Žif uncov-
ered. 60 cm. A standard glass aquarium of convenient size
can also be used and may be the cheapest solution. The
rectangular wooden boxes require less space, because they
are made so that one can be put on top of the other. In that
way, an experienced observer can record sex behavior in
six rats simultaneously without excessive head turning. It
must be noted that more complex test cages need to be
used in some studies. It is possible to make a test arena
with a glass floor fitted with a mirror inclined 458 under-
neath allowing for a ventral view, or two-level cages
where the male may be free to move around the cage, etc.
It is convenient to have the test arenas located in a quiet
room, lit with dim white light Žabout 30 lux.. Red light is
sometimes preferred. However, there is no difference in
sex behavior between rats observed under dim or bright
white light.
Standard items of sex behavior are normally recorded
with a computer keyboard. There are several programs
available for the PC of which we use two Žone described in
w9x, and another, CADA Žcomputer assisted data acquisi-
tion., available by request from Wayne A. Dornan, Dept.
of Psychology, Illinois Wesleyan University, Bloomington,
IL 61702-2900, USA.. If detailed descriptions of behav-
ioral items in addition to sex behavior are planned, the
Observer system ŽNoldus, P.O. Box 268, 6700 AG Wa-
geningen, The Netherlands, E-mail info@noldus.nl, on-line
information available on the World Wide Web,
http:rrwww.diva.nlrnoldusr. proposes expensive hard-
and software that is most convenient. Some laboratories
use in-house software, which may be available upon re-
quest. Manual data acquisition with a simple stopwatch
Žpreferably using the decimal system, i.e., minutes and
hundredths of minutes. is perfectly possible although im-
practical when several rats are observed simultaneously.
If video recordings are to be made, any video camera of
good quality is adequate. The camera can be mounted in
front of or above the test arena. A horizontal view facili-
tates observation of some behavior patterns but requires
more space.
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A. Agmor Brain Research Protocols 1 (1997) 203–209
4. Detailed procedure
4.1. Housing
A. Upon arrival to the laboratory, animals are housed in
a room with reversed lightrdark cycle in standard animal
cages, singly, in pairs or in larger groups.
4.2. Screening tests
B. The animals are transported to the observation room
and left there for 10 min or more. This may be important if
the transport involves elevators, noise or sudden bright
light. One male is then placed in each test arena Ž1–6
depending on the capacity of the observer.. About 5 min
later, the test is started by the introduction of a receptive
female in each test arena. Usually the female displays
several proceptive behaviors shortly after being introduced.
These include ear-wiggling Žrapid anteroposterior vibra-
tions of the ears., darting Ža short run where the female
abruptly stops presenting her posterior to the male. and
hopping Ža short jump with stiff legs followed by immobil-
ity and presentation.. If none of these behaviors is dis-
played, the female should immediately be replaced.
The following male behaviors are always registered:
Ž1. Mounting the female. The male normally mounts
from the rear, sometimes posing his forelegs over the
female’s back, and makes rapid Ž17–22 Hz. anteroposte-
rior pelvic thrusts for about 300 ms. He then dismounts
rather slowly. After a mount, the male frequently licks his
genital region. Mounts are performed in bouts with short
Ž5–10 s. intermount intervals. A bout is defined as a
sequence of mounts, with or without vaginal penetration,
uninterrupted by any behavior that is not oriented toward
the female except genital self-grooming. The number of
mounts within a bout varies usually between 1 and 5
Žmean about 2.. Between each bout there is a longer
Ž20–80 s. pause, during which the male may engage in
other behaviors. Before, at the beginning of, or during the
mount the female assumes a lordosis posture. If no lordosis
is displayed, the female should immediately be replaced.
An inexperienced male, and sometimes also experienced
males, may mount the female with incorrect orientation,
e.g. on the flanks, the head, etc. Such mounts are not
scored if there is no special reason to do so.
Ž2. Intromission Žvaginal penetration.. This behavior
starts with a mount, but suddenly the male makes a deep
thrust forward and stops pelvic thrusting. He then vigor-
ously withdraws and always licks his genitals. A male will
never mount again immediately after an intromission.
Ž3. Ejaculation. This behavior starts with an intromis-
sion, but after vaginal penetration Žthe deep forward thrust.
the male remains on the female for 1–3 s. Rhythmic
contractions of the posterior abdomen are clearly visible.
He then slowly raises with his forelegs held open. At
ejaculation, it is the female that moves away from the
male. He then licks his genitals and remains inactive for
205
several Ž4–7. min. In a strict sense, ejaculation is a term
referring to the expulsion of seminal liquid and not to a
behavioral pattern. However, the behavioral pattern de-
scribed is most frequently associated with actual expulsion
of semen. This latter can be verified by examining the
female’s vagina for the presence of a vaginal plug immedi-
ately after the apparent ejaculation. Rat semen coagulates
rapidly and forms a clearly visible plug. The distal part of
the vagina may be gently dilated by a forceps and the plug
extracted. It is possible to determine the spermatozoa
content w3x.
Some standard parameters of sexual behavior are regis-
tered in addition to mount, intromission and ejaculation.
Ž1. Mount latency. Time from the introduction of the
female until the first mount. Incomplete mounts Žwithout
pelvic thrusting. or badly oriented mounts are disregarded.
Ž2. Intromission latency. Time from introduction of the
female until the first intromission Žvaginal penetration..
Ž3. Ejaculation latency. Time from the first intromission
until ejaculation.
Ž4. Postejaculatory interval. Time from ejaculation until
the next intromission. Exceptionally, the first mount fol-
lowing ejaculation is taken as the endpoint of the postejac-
ulatory interval. This should be avoided, however, in order
to maintain consistency in the literature.
All these parameters are automatically calculated by the
two softwares mentioned above. Note that the CADA calls
the ejaculation latency ‘‘adjusted ejaculation latency’’.
These programs express the data in seconds. A decimal
system with the unit minute can be used instead. This
makes data presentation more economical if not too many
decimals are used.
Some additional parameters are frequently calculated.
Ž5. Intromission ratio. This is a derived parameter ob-
tained by dividing the number of intromissions by the
number of mounts plus number of intromissions.
Ž6. Inter-intromission interval ŽIII.. The ejaculation la-
tency divided by the number of intromissions.
Ž7. Copulatory rate. The number of mounts plus number
of intromissions divided by the time from the first mount
until ejaculation Žnot the ejaculation latency..
The screening tests are normally ended at the first
ejaculation Žif there is no special reason for recording the
postejaculatory interval.. If a male does not intromit within
30 min, the test is ended for that male and he is considered
sexually inactive. If a male has intromitted but not ejacu-
lated within 30 min, we give him 30 min after the first
intromission to ejaculate. Otherwise he is considered inac-
tive.
Males that are inactive at the first andror second
screening test can nevertheless be tested a second or third
time. In case that these males ejaculate at any of these
tests, they can be given additional screening tests until they
have had a total of three ejaculations. If such males are
included in experiments, the mean sexual activity will be
lower than if only males that always ejaculated are used.
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A. Agmor Brain Research Protocols 1 (1997) 203–209

Furthermore, if a repeated measure design is used, which

is frequently the case, the failure of a male to copulate at
one test renders the other tests with that male useless. This
represents a waste of time and effort that may be frustrat-
ing. I would therefore not recommend to include animals
that are irregular copulators except in the case of a parallel
group design or if there are special reasons for studying
animals with low sexual activity. There are, however, more
efficient means to obtain low-active animals Žsee Section
6..

4.3. Experimental tests

C. These tests are similar to the screening tests, with the

following exceptions:
Ž1. One or several experimental treatments are adminis-
tered before the test, in such a way that an equal number of
animals receive each treatment.
Ž2. The treatments are rotated between tests in such a
way that all animals receive all treatments. The order of
treatment can be randomized for each animal, counterbal-
anced or distributed according to a Latin square design.
Ž3. The test is ended after the first postejaculatory
interval, if the intromission latency exceeds 15 min, or if
the ejaculation latency or postejaculatory interval exceeds
30 min.
4.4. Preparation of sexually receptiÕe females
D. Ovariectomy. The female may be anesthetized with
any short-acting anesthetic. We use methohexital ŽEli Lilly
and Company, trade name Brietal in Europe and Brevital
in America. at a dose of 40 mgrkg injected intraperi-
toneally. With the female in a prone position, a medial
dorsal incision, about 1.5 cm long, is made through the
skin midway between the last rib and the knee. The skin is
then pulled about 1 cm to the left, and a second incision is
made through the muscle layer and into the peritoneal
cavity. If this incision is correctly placed, the ovary will be
located immediately underneath, invisible in the midst of a
mass of fat. The fat is withdrawn, the ovary is separated
and the oviduct ligated with catgut or silk Žsee Fig. 1.. The
ovary is cut away and the incision sutured. The ovary on
the opposite side is then removed through a separate
incision. The entire operation does not require more than
10 min when some practice has been obtained.
E. Preparation of steroids. Independently of whether
intact or ovariectomized females are to be used, female
sexual behavior needs to be activated by ovarian hor-
mones. In principle, it would be possible to use normally
cycling females at the moment of proestrus–estrus. In
order to determine this moment, vaginal smears need to be
taken and examined, which requires some time. Further-
more, naturally cycling females get frequently pregnant.
Such complications are the reasons why hormone-treated
females almost always are used.
Fig. 1. Left: the approximate site of the medial dorsal incision for
ovariectomy is shown. Right, upper drawing: an incision has been made
through the muscle layer and the ovary, partially liberated from surround-
ing fat, is visible as a dark sphere adjacent to the oviduct. In the lower
drawing the oviduct has been ligated and the ovary can now be removed.
For further details, see text.
Oil is added to crystalline estradiol benzoate and the
mixture is heated to 608C for at least 1 h and then
thoroughly shaken. Progesterone is prepared in the same
way. The final concentration should be 100 m grml of
estradiol benzoate and 5 mgrml of progesterone. Estradiol
benzoate is injected about 52 h before the females are to
be used and progesterone about 4 h before either to intact
or ovariectomized females. Both hormones are injected
subcutaneously in a volume of 0.2 mlrrat on the back or
flanks. The doses are 20 m grrat for estradiol benzoate and
1 mgrrat for progesterone. This treatment assures intense
proceptivity and receptivity. Females can be used once a
week for several months. In our experience, ovariec-
tomized females show a higher degree of receptivity than
intact ones. This affirmation, however, is not based upon
quantitative comparisons.
Instead of injecting estradiol benzoate, a 5 mm long
Silastic capsule Ž1.57 mm i.d. and 3.18 mm o.d., Dow
Corning Medical Corporation. containing crystalline estra-
diol can be subcutaneously implanted at the moment of
ovariectomy or anytime thereafter. A detailed description
of the procedure for making silicone capsules has been
published w11x. Progesterone, 1 mgrrat, is injected about 4
h before test. With this technique, females can be used
twice or more per week during at least 2 months.
5. Results
The mount latency is a measure of sexual motivation
w16x. If only males that ejaculated at each of the screening
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A. Agmor Brain Research Protocols 1 (1997) 203–209
tests are used, it should normally be under 1 min and only
exceptionally above 5 min. The intromission latency is
frequently the same as the mount latency Žthe first mount
ending with intromission., but it may also be somewhat
longer. Intromission requires penile erection and coordi-
nated activity of the striated penile muscles and is there-
fore not entirely determined by sexual motivation. The
ejaculation latency, again in vigorous copulators, varies
normally between 4 and 10 min although values outside
this range are not uncommon. The postejaculatory interval
varies usually between 4 and 7 min. In fact, the interindi-
vidual variation in postejaculatory interval is considerably
less than in mount, intromission or ejaculation latency.
Although once considered to be a measure of sexual
motivation, it varies rather independently of the mount
latency and responds differently to several experimental
manipulations. The interpretation of this interval is, there-
fore, unclear.
When a rat fails to copulate, no latencies or postejacula-
tory interval can be recorded. If a repeated measures
design is used, this obliges to drop all data from that
particular subject. To avoid this, latencies are sometimes
assigned to non-copulating animals. In this case, the cut-off
time is the value assigned.
The number of mounts is quite variable, ranging be-
tween 0 and 20 or more. This number probably reflects
sexual motivation, but may be confounded with other
factors, and should be interpreted with caution. The num-
ber of intromissions, usually between 5 and 10, shows the
ease by which ejaculatory reflexes are activated. The intro-
mission ratio varies between 1 Ža male making only intro-
missions. and 0 Ža male making only mounts. with a
normal range of 0.5–0.7. This parameter represents the
potency, in other words the efficiency of erection and
penile orientation.
The inter-intromission interval and copulatory rate may
reflect a mixture of sexual motivation and potency, and it
is not clear that these parameters provide any useful
additional information.
Most male rats are able to make between 5 and 8
ejaculations in a series. A postejaculatory interval of more
than 90 min indicates that the male is sexually exhausted,
and the intensity of sexual behavior will be reduced during
the following 2 weeks. In order to be able to use the males
frequently, sex behavior tests are therefore usually ended
after the first postejaculatory interval. Exceptionally it can
be of interest to study the evolution of behavior during
successive ejaculations andror the number of ejaculations
achieved before exhaustion. In that case, it will be found
that the number of mounts and intromissions as well as the
ejaculation latency are reduced for the second ejaculation.
This reduction persists at the third and frequently at the
fourth ejaculation. Thereafter these parameters increase
above what was found at the first ejaculation. The poste-
jaculatory interval shows an exponential increase from the
second ejaculation and onwards.
207
6. Discussion
6.1. Troubleshooting
The most serious problem that may be encountered is
that a group of males displays an unusually low level of
sexual activity, with several males failing to copulate at
several screening tests and others having long latencies.
There is neither a simple explanation nor a simple solution
to this problem. Some males become sexually activated by
a change of female, and this can be tried at regular
intervals. Low female proceptivity may be a reason for low
male sexual activity, particularly when the male has no
previous sexual experience. It is recommended to carefully
check female behavior, and if necessary to replace the
females. Inconvenient animal housing Žinsufficient ventila-
tion, inadequate humidity or temperature, dirty bedding
material. may also adversely affect sexual behavior, but
such events are both inadmissible and rare in most animal
facilities. The observation room, however, may become
too hot if not correctly ventilated.
Occasionally males Žor females. may show a consider-
able amount of aggression towards their partner. A change
of female may solve this problem.
6.2. AlternatiÕe and support protocols
Male sexual behavior requires normal functioning of the
hypothalamic-pituitary-testicular axis. Treatments that
modify the concentration of circulating sex hormones may
therefore modify sex behavior. Reduced hormone levels
have no immediate effect, because sexual behavior begins
to decline about 1 week after castration and does not
disappear until 3 weeks after castration. However, in-
creased concentration of circulating testosterone has been
reported to modify sexual behavior within 1 h w12x. Experi-
mental manipulations that alter the endocrine system can,
therefore, erroneously be interpreted as directly affecting
mechanisms controlling sexual behavior. In order to avoid
this kind of confusion, castrated animals equipped with a
constant release source of gonadal hormones can be used.
A Silastic capsule, similar to the one described for use in
females but 20–40 mm long and filled with crystalline
testosterone, assures serum testosterone concentrations
within the physiological range for at least 3 months w5x.
In some circumstances, behavioral effects of brain le-
sions or drug treatments are only detectable in rats with a
subnormal sexual activity w1,2x. It is sometimes argued that
stimulatory effects are more easy to detect in animals with
low sexual activity, although there is no empirical evi-
dence in favor of this notion. Animals with low sexual
activity are obtained by treating castrated males with low
doses of testosterone. Either short Silastic capsules Ž5 mm.
or weekly injections of testosterone propionate Žwith 0.4
mgrkg about 50% of the males display intromission and
with 0.2 mgrkg intromissions are almost absent but some
208 ˚
A. Agmor Brain Research Protocols 1 (1997) 203–209
mounting behavior remains. can be used for this purpose.
It is also possible to select intact males with irregular and
low sexual activity, but it is often difficult to obtain a
sufficient number. Furthermore, it is not evident that such
animals are representative.
Sometimes it is useful to record several behaviors other
than copulatory acts. This is particularly useful in studies
of drug action w1,14x and the effects of brain lesions w15x.
The analysis of exploratory behaviors Žrearing, sniffing,
gross motor activity. may provide useful information about
the animals’ general arousal and makes it possible to
determine if treatment effects are specific to sexual behav-
ior. Items of non-sexual social interaction can also be
recorded. Among these are sniffing or grooming the part-
ner, crawling under or over the partner and genital explo-
ration. The intensity of these behaviors is not different
between rats that copulate and rats rendered sexually inac-
tive by castration or brain lesion w15x making their associa-
tion to sexual behavior doubtful. On the contrary, pursuit
of the female Žthe male running behind her in close
contact. is a good indicator of imminent copulation w6,15x.
If a treatment effect is specific to sexual behavior, only
pursuit of the female should be modified. Otherwise, a
non-specific effect on social interactions is probably at
hand. This can be further confirmed in tests where the
receptive female is replaced by a castrated male. In the
latter case, no or very little copulatory behavior or pursuit
is observed. If a treatment modifies interactions with a
castrated male, its effects are not specific to sexual behav-
ior.
An elegant way to distinguish sexual motivation from
sexual performance is to make access to the female contin-
gent on an operant response. Male rats may be trained to
lever-press in a Skinner box modified in such a way that
the lever activates a mechanism that introduces a receptive
female into the box w7x. The training is rather complex, but
the extra effort required is justified in studies where analy-
sis of sexual motivation is the main objective. Another
original way to study drug effects on sexual motivation is
to allow the males to copulate until exhaustion and then
determine how rapidly they recuperate sexual behavior
w17x.
Occasionally treatment effects become particularly evi-
dent if females with low proceptivity are used. Moreover,
it seems that many treatments reducing general arousal
have no effect on sexual behavior when intensely procep-
tive females are used, but are deleterious when female
proceptivity is low. One easy way of reducing or entirely
eliminating female proceptivity without altering hormone
treatments Žwhich by itself could affect male behavior. is
to administer a dopamine antagonist to the female shortly
before the test. We use cisŽ Z .-flupenthixol ŽLundbeck,
Copenhagen, Denmark. injected intraperitoneally 30 min
before the test. A dose of 1 mgrkg much reduces procep-
tivity while 2 mgrkg produces an essentially immobile but
perfectly receptive female.
It may also be desirable to study motor activities spe-
cific to copulation. The most evident is perhaps the pelvic
thrusting associated with mounts and intromissions. The
purpose of this thrusting is to bring the penis in contact
with the vaginal orifice. It also activates or strengthens the
female’s lordosis. There exists an accelerometric technique
for the analysis of frequency, duration and amplitude of
pelvic thrusting w4x. Little is known about effects of drugs
and lesions on this behavior.
Intromission is not possible without adequate erection
and coordinated activity in the penile muscles. There exists
a technique for recording, by telemetry and in copula, the
pressure of the corpora cavernosa w8,18x. It is also possible
to obtain electromyograms from the striated penile muscles
during copulation w10x. If the behavioral data show an
effect on intromission ratio, it should always be suspected
that some penile activity has been modified. The above-
mentioned techniques may be used to determine whether
this is the case.
Male rats ‘‘sing’’ before, during and after copulation
w20x. The female is also singing, hopefully in beautiful
concert with the male. Very little is known about the role
and the physiological control of these ultrasound vocaliza-
tions, but they constitute undoubtedly an element of sexual
behavior.
7. Quick procedure
As a specific example, I describe the procedure for
testing the effects of drugs on sexual behavior. With minor
modifications, this procedure can be applied to most kinds
of studies of sex behavior.
A. Housing the rats under a reversed lightrdark cycle
for about 3 weeks.
B. While the males are habituating to the lightrdark
cycle, females to be used as mating partners may be
ovariectomized.
C. The males are subjected to three screening tests with
receptive females in order to assure that experimental
males are sexually active and have acquired some experi-
ence. The tests are separated by at least 72 h and are
performed during the dark phase of the lightrdark cycle.
Ideally, only males ejaculating at each test are used in
experiments.
D. Experimental tests are begun and can normally be
performed 3 times a week until the experiment is com-
pleted.
E. Recording of behavioral parameters can be made
either directly during copulation or later from video tape.
If possible, the observer should be blind to experimental
treatments.
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A. Agmor Brain Research Protocols 1 (1997) 203–209
8. Essential literature references
Original papers: ref. w6x.
Textbook chapters: ref. w13x.
Review papers: ref. w16x.
References
˚
w1x Agmo, A. and Picker, Z., Catecholamines and the initiation of
sexual behavior in male rats without sexual experience, Pharmacol.
Biochem. BehaÕ., 35 Ž1990. 327–334.
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