ESTROUS CYCLE

The estrous cycle comprises the recurring physiologic changes that are induced by reproductive hormones in most mammalian
placental females. Humans undergo a menstrual cycle instead. Estrous cycles start after puberty in sexually mature females and are
interrupted by anestrous phases or pregnancies. Typically estrous cycles continue until death. Some animals may display bloody
vaginal discharge, often mistaken for menstruation, also called a "period".

Stages of the canine estrous cycle can be defined by sexual behavior, physical signs (vulvar swelling, vaginal bleeding) or by vaginal
cytology. The period of receptivity to a male varies considerably among bitches; some bitches are receptive well before and after the
period of potential fertility. Similarly, signs such as "proestrus bleeding" are often unreliable indicators; some bitches bleed very little
and other show bleeding through estrus and into diestrus.

Since cytologic changes reflect the underlying endocrine events of the cycle, they are almost always a better predictor of the "fertile
time" and gestation length than are behavioral or physical signs.

Cytologic changes through the canine estrous cycle reflect changes in blood concentrations of estrogen. As depicted below, and
described in more detail in the section on Canine Reproduction, estrogen levels rise prior to and during proestrus and fall in
conjunction with the preovulatory surge of luteinizing hormone. Rising levels of estrogen induce the "cornification" that is
characteristic of smears examined during estrus. Ovulation occurs two days after the LH surge.

Anestrus
Anestrus refers to the phase when the sexual cycle rests. This is typically a seasonal event and controlled by light exposure through the
pineal gland that releases melatonin. Melatonin may repress stimulation of reproduction in long-day breeders and stimulate
reproduction in short-day breeders. Melatonin is thought to act by regulating hypothalamic pulse activity of gonadotropin-releasing
hormone. Anestrus is induced by time of year, pregnancy, lactation, significant illness, chronic energy deficit, and possibly age.

Intermediate and parabasal cells predominate in

smears taken during anestrus. Superficial cells are
absent or found in very small numbers.
Neutrophils may also be present or absent.

Proestrus
One or several follicles of the ovary are starting to grow. Their number is specific for the species. Typically this phase can last as little
as one day or as long as 3 weeks, depending on the species. Under the influence of estrogen the lining in the uterus (endometrium)
starts to develop. Some animals may experience vaginal secretions that could be bloody. The female is not yet sexually receptive.

Serum concentrations of estrogen rise during proestrus, leading to capillary breakage and leakage of red blood cells through uterine
epithelium, as well as proliferation of the vaginal epithelium.

Examination of vaginal smears from early to late proestrus will reveal a gradual shift from intermediate and parabasal cells to
superficial cells. Typically, red blood cells are present in large numbers and neutrophils are commonly observed. Large numbers of
bacteria are also often present.

In some bitches, proestrus can persist for two to three weeks. In such cases, prolonged lack of receptivity may suggest the need to
artificially inseminate or force-breed the animal. Examining vaginal smears in such cases will alleviate such concerns - certainly, if
more than a very small percentage of cells are parabasals and s mall intermediates, breeding is a waste of time.
 Superficial cells increases and also erythrocytes
increases

Estrus
Estrus refers to the phase when the female is sexually receptive ("in heat," or "on heat"). Under regulation by gonadotropic hormones,
ovarian follicles are maturing and estrogen secretions exert their biggest influence. She then exhibits a sexually receptive behavior, a
situation that may be signaled by visible physiologic changes. A signal trait of estrus is the lordosis reflex, in which the animal
spontaneously elevates her hindquarters.

In some species, the vulvae are reddened. Ovulation may occur spontaneously in some species (e.g. cow), while in others it is induced
by copulation (e.g. cat). If there is no copulation in an induced ovulator, estrus may continue for many days, followed by 'interestrus,'
and the estrus phase starts again until copulation and ovulation occur.

The defining characteristic of cytologic estrus is the predominance of superficial cells. Most, but not all, bitches will undergo full
cornification, and the smear will reveal a monotonous pattern composed almost exclusively of anucleate superficial cells.

If the bitch has been bred within a day of preparing a vaginal smear, it is quite likely that sperm will be observed among the epithelial
cells. Indeed, careful examination for sperm in a smear taken within a few hours of an alleged breeding is a fairly reliable means of
confirming or denying such an incident. In the image above, an intact sperm (left panel) and a sperm head (right panel) are present
next to superficial cells.

Diestrus
Diestrus is characterised by the activity of the corpus luteum that produces progesterone. In the absence of pregnancy the diestrus
phase (also termed pseudo-pregnancy) terminates with the regression of the corpus luteum. The lining in the uterus is not shed, but
will be reorganised for the next cycle.

The onset of diestrus is marked by a precipitous decline in the number of superficial cells and reappearance of intermediate and
parabasal cells. Most commonly, the cellular profile changes within a single day from essentially 100% superficial cells to less than
20% superficial cells. However, it is best to confirm the onset of diestrus by examining a smear prepared on diestrus day 2.
The significance of identifying the onset of diestrus is that it is a considerably more accurate predictor of the time of ovulation, and
hence gestation length, than sexual behavior.

Dogs ovulate 5-7 days prior to the onset of diestrus (7-9 days after the preovulatory LH surge), and hence, gestation length is usually
57 + 1 day from the onset of diestrus day 1. The period of behavioral estrus is variable, and often extends up to several days before
and/or after cytologic estrus. Gestation lengths calculated from the onset or cessation of receptivity are correspondingly inaccurate.
The onset of diestrus also correlates well with loss of fertility, and breedings after the diestrus shift are rarely fertile.

Metestrus
During this phase, the signs of estrogen stimulation subside and the corpus luteum starts to form. The uterine lining begins to secrete
small amounts of progesterone. This phase typically is brief and may last 1 to 5 days. In some animals bleeding may be noted due to
declining estrogen levels.

The period of metestrus begins with the cessation of estrus and laster for about 3 days. Primarily, it is period of formation of the
corpus luteum (corpora lutea with multiple ovulation). However, ovulation occurs during this period in cows and dies. Also, a
phenomenon known as metestrus bleeding occurs in cows. During late prorestrus and estrus, high estrogen concentrations increase the
vascularity of the endometrium, this vascularity reaching its peak about 1 day after the end of estrus. With decline estrogen levels,
some breakage of capillaries may occur, resulting in a small loss of blood. This will be noticed as a patch of blood o the tail
approximately 35 to 45 hours after the end of estrus. It is not an indication of conception or a failure to conceive. Also, it should not be
confused with menstrual bleeding, which occurs in humans.

Don Velasquez
Seat #35
Histology Lec Assignment