Genital Tract Motility in the Domestic Hen

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B. ALEXANDER AND A. WOLFSON nese quail, (Coturnix coturnix japonica). Science, 158: 1478-1479. Tanaka, K., F. B. Mather, W. O. Wilson and L. Z. McFarland, 1965. Effect of photoperiods on early growth of gonads and on potency of gonadotropins of the anterior pituitary in Coturnix. Poultry Sci. 44: 662-665. van Tienhoven, A., 1961. Endocrinology of reproduction in bird. In Sex and Internal Secretions. Ed. W. C. Young, Williams and Wilkins Co., N.Y., pp. 1088-1169. Voogt, J. L., J. A. Clemens and J. Meites, 1969. Stimulation of pituitary FSH release in immature female rats by prolactin implants in the median eminence. Neuroendocrinology, 4 : 157163.

Genital Tract Motility in the Domestic Hen T. W. CHEN* AND R. O. HAWES Department of Animal Science, Macdonald College (McGill University), Que., Canada (Received for publication October 27, 1969)

T

HE role of the avian oviduct in egg formation has received attention for many years. More recently, the oviduct's role in the storage and transport of sperm cells has been investigated (see reviews by Hafez, 1968; and Van Teinhoven, 1968). While the spontaneous motility of the oviduct is necessary for both the transport of ovum and of spermatozoa, there has been a minimum of work in the area of oviduct motility. The spontaneous motility pattern of avian vaginal and uterine muscle strips was studied in vitro by Sykes (1955). He demonstrated that contractions of the vagina differed from those of the uterus, being of smaller amplitude and greater frequency. No spontaneous motility was

* Present address: Department of Biology, Nanyang University, Singapore 22 Contribution from the Faculty of Agriculture, McGill University, Macdonald College, Que., Canada. Journal Series No. 604.

found in the vagina of a non-laying hen and the amplitude of uterine contractions was much lower than those of a laying hen. Sykes was unable to demonstrate differences between regions within the same part of the tract. The immature chicken responds to an exogenous source of estrogen by a significant increase in its oviduct size and the size of the surrounding ligaments (Brant and Nalbandov, 1956). The immature mammalian uterus displays no spontaneous activity, treatment with estrogen induces drastic changes in membrane and myoplasmic structures and it thus becomes spontaneously active (Csapo, 1961). The present study was an attempt to determine the spontaneous motility patterns of the hen's oviduct at various ages, to study the effect of exogenous estrogen on the oviducal motility in immature chickens and to determine the motility patterns of circular muscles from various areas of the mature oviduct.

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tin injection by an objective and quantitative method. Endocrinology, 80: 641-655. Riddle, O., and R. W. Bates, 1933. Concerning anterior pituitary hormones. Endocrinology, 17: 689-698. Riddle, O., and P. F. Braucher, 1931. Studies on the physiology of reproduction in birds. XXX. Control of the special secretion of the crop gland in pigeons by an anterior pituitary hormone. Am. J. Physiol. 97: 617-625. Saeki, Y., and Y. Tanabe, 1955. Changes in prolactin content of fowl pituitary during broody periods and some experiments on the induction of broodiness. Poultry Sci. 34: 909-919. Sayler, A., and A. Wolfson, 1967. The avian pineal gland: A progonadotropic response in the Japa-

GENITAL TRACT MOTILITY

MATERIALS AND METHODS

2) Removal and isolation of tissues: After recording live body weight, birds were killed by dislocating the cervical vertebrae near the atlas. The ovary and oviduct were removed, excess connective tissue and fat trimmed from them and their weights recorded. The tissues were then placed in Krebs solution maintained at 41 °C. The entire magnum and uterus were used for recording the longitudinal muscle motility of 4-16 week old birds. For the 20 and 24 week old group the entire uterus was used but only a section of the midmagnum (4 cm.). In addition, the tracts of sexually mature birds were divided into 31 regions and transverse sections (1 cm. wide) were removed for circular muscle recordings. Samples were taken from the infundibulum, magnum, isthmus and vagina, and opened on the ventral side to form a strip. For uterine recordings a cross section of 1 cm. wide and 3-3^ cms. long was removed for circular muscle activity. Recording equipment and procedures: The basic unit of the Grass Model 7 Polygraph

used for the present work consisted of a console, equipped with a chart driver system, a power supply, a writer unit, 4 D.C. driver amplifiers and 4/7P1 preamplifiers. Recordings were made in curvilinear coordinates. The apparatus designed for studying the motility of circular and longitudinal muscles of the genital tract consisted of the following parts: 1. Grass force-displacement transducer (FT03C). 2. Rubber stopper. 3. Glass tubes (2.5 X 20.S cm.). 4. Temperature control unit (Precision Scientific Co.). 5. Water bath. 6. Flask containing Krebs solution. 7. Outlet tube for rinsing. 8. Oxygen carrying tube. For recording muscle motility, the upper end of the tissue was suspended from the force-displacement transducer with a thread while its lower end was firmly fixed to a hook on the rubber stopper at the lower end of the glass tube. Forty cc. of Krebs solution (41 °C.) was then added to the glass tube and 0 2 (95%) + C0 2 (5%) were allowed to bubble through a hypodermic needle at the bottom of the tube. The glass tube was then immersed in the water bath (41°C.) to a level equal to that of the Krebs solution in the tube. The sensitivity of the preamplifier was calibrated in such a way that when at 1 mv. /cm., one gram of tension applied at the transducer caused a 2.5 cm. pen deflection on the recording chart. Recordings were made after an acclimatization period of 10-15 minutes. For analyzing tract motility a standard portion (10 min. of recording at a chart speed of 0.5 mm./min.) of the tracing was taken and the average amplitude (M) and average frequency (F) of the contractions were measured. From these values, an index of activity (I) was calculated according to the equation I = F XM. All reported values are based on a mean of 3 birds.

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1) Animals: Female Rhode Island Reds (4, 8, 12, 16, 20, 24 weeks old) were used. Chicks were hatched in March-April, brooded under conventional conditions from 1 to 8 weeks, moved to "growing shelters" for 8-20 weeks of age, and to a windowless laying house for the remainder of the experimental work. Mature birds destined for experimental use were kept in cages for several weeks although they had previously been in floor pens. Birds of all age groups received commercial rations and water ad libitum. All mature birds received 14 hours of artificial light. In addition, 4 mg./day of diethylstilbestrol dipropionate was injected im. for 6 days to 8, 12, and 16 week old birds, to study the effect of exogenous estrogen.

641

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T. W. CHEN AND R. O. HAWES

age (0.77% and 0.07%, respectively) but increased considerably after 16 weeks. At 24 weeks of age, the percent body weight of the genital tract and ovary reached 2.75% and 2.39%, respectively. These values were relatively high when compared with those of the Fayoumi breed of the same age (Hafez, 1955).

'*

'«

20

24 WEEKS

FIG. 1. The effect of age on the growth of the ovary, genital tract and the motility of the magnum.

RESULTS AND DISCUSSION

The growth of the genital tract. The changes in weights of the genital tract and ovary with age, are seen graphically in Figs. 1 and 2. The slopes of the lines representing these two factors were relatively low until the period between 6 and 20 weeks of age when the slopes began to increase greatly. In birds, the ovary and oviduct were classified by Hafez (19SS) as late maturing organs. The proportion of genital organs to body weight was negligible at hatching and up to the age of six months in Fayoumi chickens. Similar results were obtained from Rhode Island Reds in the present study where the growth of the ovary and oviduct were relatively slow from 4 to 12 weeks of age. The weights were 1.18 ± 0.12 g. and 1.24 ± 0.52 g. for ovary and tract respectively at the latter age. At 24 weeks, the weights were S1.8S db 9.25 and 59.70 ± 0.90 g., respectively. It was found that the percent body weights of the genital tract and of the ovary were relatively low up to 16 weeks of

4

•

12

14

20

24 WEEKS

FIG. 2. The effect of age on the growth of the ovary, genital tract and the motility of the uterus.

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•

The effect of age upon spontaneous motility of the genital tract. The spontaneous motility of the magnum varied with age both in magnitude and frequency of contraction (Fig. 1). The motility pattern recored at 4 weeks showed rapid (20.5 ± 0.41/min.) but irregular contractions of low magnitude (0.005 ± 0.001 g.). Magnitude of contractions were slightly increased to 8 weeks of age with little change to 12, and thereafter increased sharply to a maximum of 0.78 g. at 24 weeks. Frequency was maximum at 8 weeks (23.80 con./min.) and declined slowly to 12 weeks, then sharply to 20 weeks (9.70 con./min.) with little change to 24 weeks (Figs. 1,3). Spontaneous activity of the longitudinal

GENITAL TRACT MOTILITY

643

WEEKS

MAGNUM

UTERUS

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muscles of the uterus also varied with the biologically active estrogen in the blood of age of the bird (Fig. 2). The frequency and 16 to 28 day old pullets. Another possibilmagnitude of contractions of the uteri from ity which might cause a difference in geni4 week old birds were 7.40 ± 2.70/min. tal tract activity in young animals would and 0.08 ± 0.01 g., respectively. These be the sensitivity of the recording technivalues were relatively low compared to que. Since the genital tract of 4 week old those of the older birds. The magnitude re- birds is very small (0.02 g.) and of very mained quite similar for the 8 and 12 week weak musculature, it is expected that its old groups but showed a marked increase contractions would be of very small magniat 16 weeks continued this trend through tude and require high amplification before 20 and 24 weeks of age (1.41 ± 0.42 g. at they could be recorded. Ovarian weight, and tract weight were 24 weeks). Frequency increased to a peak at 16 weeks (26.90 ± 0.90/min.) then de- increased as the birds aged from 4 to 24 creased at 20 weeks with little change at weeks. These changes were shown to be as24 weeks (12.17 ± 1.23/min.) (Fig. 2, 3). sociated with the changes in the frequency, The above results show that spontaneous magnitude and consequently the index of motility of the genital tract occurred as activity (Figs. 1, 2). early as 4 weeks of age in Rhode Island Correlation coefficients calculated for the Reds. This finding tends to agree with that factors which influence spontaneous motilof Chen (1964) who recorded in situ and ity are shown in Table 1. The relationship in vitro, the spontaneous genital tract mo- between the increase in body weight and tility in young rabbits and rats. The results the development of ovary and genital tract were, however, in contrast to those of Far- was positively related with the magnitude ner (1962) who showed no spontaneous of contraction and the index of activity. contractions in young rats of 4 months old. The correlations between frequency of conIt may be that the estrogen, which is re- traction and body, ovarian and tract weights. sponsible for the growth and activity of the were generally negative and in no case sigtract, was relatively low in young mammals nificant. compared to avians. Kornfeld and Nalbandov (1954) demonstrated the presence of a The effect of exogenous estrogen. Intramuscular injections (4 mg./day for 6 days) of stilbestrol caused an increase in the weights of genital tracts from 8, 12 and 16 week old ^Y^fV't'M'w^;/ -^ —— birds of 350, 808, 868-fold respectively h^^^AOt^S^E over the controls of the same age (Fig. 4). 8 ;./"#,(^«* There was a reduction of about one-fold in the ovarian weights in the 16 week old birds, as compared to that of the untreated 16" 'l****W**~**,r'**"«'*''>"W birds. Similar results were obtained from the works of Juhn and Gustavson (1930), Kar (1947) and Brant and Nalbandov (1956). The increase in the weight of the estrogen-induced oviduct was attributed by FIG. 3. Representative tracings of the sponta- several workers to the increase in the total neous genital tract motility in birds of different dry matter, protein and nucleic acid content (Mueller, 1957; Brown and Jackson, ages.

644

T. W. CHEN AND R. O. HAWES TABLE 1.—Correlations for factors involved in influencing spontaneous motility Body wt. Ovary Wt. Tract Wt.

1 Mag. F. 1 Ut. 1 Mag. M. | Ut. 1 Mag. Ut.

0.58 0.71 -0.35 0.34

Tract wt.

F. Mag.

M. Ut.

Mag.

Ut.

0.96** -0.27 -0.28

-0.33 -0.35

0.84* 0.80*

0.87* 0.97**

0.96** 0.98**

-0.37

0.87* 0.76*

0.81* 0.92**

0.92** 0.90*

-0.37

0.13 0.98* 0.30

0.96*

* Significant at 0.05. ** Significant at 0.01. F.—Frequency of Contraction. M.—Magnitude of Contraction. I.—Index of Activity.

1960) and to the decrease in hydroxyproline content (Anastassiadis et al., 1955). Following the induced development of the genital tract, there was an increase in the magnitude and a decrease in the frequency of the contractions in both magnum and uterus (Fig. 4). The index of activity, however, was increased in all 8, 12 and 16 week old groups compared to the controls. It was estimated, from the index of activity, that the activity of the estrogen-dominated genital tracts from 8, 12 and 16 week old birds was comparable to those of the mature or nearly mature birds. It was also noted that the uterine activity was affected much more than that of the magnal regions, showing that the musculature of the uterus was more sensitive to this particular hormone. Although the increase in the motility of the genital tract may be attributed mainly to the increase in the protein and dry matter content of the tract by estrogen treatment (Brown and Jackson, 1960), simultaneous changes in other factors caused by the treatment should also be considered. It was found that the high energy phosphate compound creatine phosphate disappeared, and the ATP concentration decreased by

50% after ovariectomy, whereas the original level was restored within about 2 days of estrogen treatment (Csapo, 1955). There has been evidence to show that estrogen treatment induced drastic changes in membrane and myoplasmic properties in the mammalian uterus. The working capacity of uterine muscle was determined by the actomyosin content of the myofibrils whose synthesis was controlled by estrogen (Csapo and Corner, 1953). Spontaneous motility of the circular muscles. In order to understand the mode of contraction in each region of the genital tract, the oviduct from 24 week old birds was divided, from the fimbria to the lower portion of vagina, into 31 regions and their motility patterns were recorded (Figs. 5, 6). The numerical value for regions in terms of frequency and magnitude of contractions is shown in Table 2. The motility pattern of the fimbria (Fig. 5A) was characterized by a series of 14-15 strong contractions with a mean magnitude of 3.19 g. at a frequency of 7/min. This train of contractions was followed by a series of weaker ones (0.78 g.) at a frequency of 3.7/min. and lasted for 7-8 min-

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I.

Ovary wt.

645

GENITAL TRACT MOTILITY

r

iimimMtoii^MiiM iM 1 JUUNlAMMK'^AxNkl^

M T",1w*,,*''W"'?-*',W»ft^^ 0 J"-•"*"! rum-la P

I B gsiiwm C D CONTROL

t

v - • *- ;

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FIG. 5. Motility patterns of circular muscles A-C D-L M N-0 P

FIG. 4. The effect of exogenous estrogen on the body weight, ovary weight, tract weight and spontaneous motility of the magnum and uterus.

••

|/MltfW^^WvMANV.

Infundibulum Magnum Magnal-isthmal junction Isthmus Isthmal-Utero Junction

were not in a regular sequence. The similarity found in the motility patterns of the infundibulum and fimbria were expected, due to the fact that there is no distinct difference in musculature of the two regions, except the relative thinness of the latter (Surface, 1912). It appeared that the mode of contraction of a particular region was highly related to its musculature construction. As the thin walled infundibulum changed to the thicker walled magnum; the proximal, middle and distal upper magnum (Fig. 5D, E, F, and Table 2) were characterized by strong rhythmic contractions of low frequency. There seemed to be little difference in the contractility pattern between the proximal and middle upper magnum, except that the motility pattern showed by the latter

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utes, after which the strong contractions resumed, thus producing an average frequency of 5.5/min. This pattern of contraction was occasionally separated (at intervals of about 10-15 min.) by a period of complete cessation of contractions which lasted for about 10 minutes. The proximal region of the infundibulum (Fig. 5B) showed rhythmic contractions of higher frequency (7.0/min.) and at an average magnitude of 1.78 g. Cyclic changes in the magnitude of contractions were noted; trains of strong contractions were followed by a series of weaker ones; the cycle repeated itself once every 4-5 minutes. The distal infundibulum (Fig. 6C) showed a similar pattern of magnitude and frequency, although the contractions

646

8

T. W. CHEN AND R. O. HAWES

J v*********^^

m *\

" iWJJJMWJMMUim FIG. 6. Motility patterns of circular muscles A-G H-I J-0

Uterus Utero-vaginal junction Vagina

TABLE 2.—Comparison of spontaneous motility of circular muscles from different regions of the genital tract Regions of tract

Frequency of Magnitude of contraction contraction con./min.) (g.)

(

was not as regular. The distal upper magnum, however, showed regular strong contractions (3.6 g.) at a frequency of 4.0/ min. Each contraction was separated from the other by a short (4-5 sec.) relaxation period. The general consistency in the motility pattern of strips from the same region provided good evidence that the tissues were able to maintain their characteristic mode of contraction under the experimental conditions. The motility pattern of the middle portions of the magnum was somewhat similar to the upper parts, except that the frequency of contractions was reduced and the magnitude slightly increased (Fig. 5G,

Fimbria Prox. Infund. Dist. Infund. Prox. Up. Magnum Mid. Up. Magnum Dist. Up. Magnum Prox. Mid. Magnum Mid. Mid. Magnum Dist. Mid. M a g n u m Prox. Low. M a g n u m Mid. Low. Magnum Dist. Low. M a g n u m Mag.-Isth. Junction Prox. I s t h m u s Dist. Tsthmus Istli.-Ut. Junction Prox. Up. Uterus Dist. Up. Uterus Prox. Mid. Uterus Dist. Mid. Uterus Prox. Low. Uterus Mid. Low Uterus Dist. Low Uterus Prox. Ut.-Vag. June. Dist. Ut.-Vag. June. Prox. Up. Vagina Dist. Up. Vagina Prox. Mid. Vagina Dist. Mid. Vagina Prox. Low. Vagina Dist. Low. Vagina

5.50 7.00 7.00 4.00 3.50 4.00 2.50 2.50 2.50 3.00 3.75 7.00 9.00 9.50 10.50 5.75 17.00 14.00 13.00 13.50 10.50 15.50 12.50 9.00 5.50 6.00 4.00 4.00 2.50 5.50 1.50

1 .98 1.78 1.75 3.50 2.91 3.60 3.15 3.58 3.57 2.13 1.07 1.41 0.59 0.69 0.40 0.47 0.46 0.58 0.95 0.38 0.44 0.26 0.15 0.11 0.20 0.56 0.90 0.43 1.52 0.52 0.21

Index of activity (10 min.) 126.33 125,00 123.00 140.00 102,00 144.25 78.75 89.50 89.25 64.00 80.00 99.50 51.75 65.75 42.70 26.75 74.25 80.50 124.75 51.50 46.75 40.75 19.25 5.79 11.75 33.80 36.00 17.05 38.0(1 21.25 3.06

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•»M.i..».i.Ji.iriVi« mi nwii.,,!,,,.,-

H, I and Table 2). The relaxation period between each contraction was increased to about 10 seconds. The frequency and magnitude of contractions of the proximal and middle portions of lower magnum differed only slightly from those of the middle regions (Fig. 5J, K and Table 2). There was, however, a distinct increase in the frequency of contraction in the distal region of the lower magnum (Fig. 5L). Since the mode of contraction in this region resembled more that of the magnal-isthmal junction (Fig. 5M) and isthmus (Fig. 5N, 0 ) , than that of the other regions of magnum, it appeared that the change in the mode of motility from magnum to isthmus began at this region. The magnal-isthmal junction (Fig. 5M) represents a transition zone in which the magnum narrows down to the isthmus. The frequency was further increased and magnitude of contraction decreased in this region with the appearance of cyclic changes in the magnitude.

GENITAL TRACT MOTILITY

Polge (1951) obtained no fertility following intra-vaginal insemination with semen containing 15% glycerol, while Allen and Bobr (1955) obtained low fertility when semen plus glycerol was deposited directly in the uterus. Moreover, Allen and Grigg (1957) found that the number of sperms reaching the infundibulum was much smaller after intra-vaginal than after intra-uterine insemination. The possibility that the uterovaginal junction may act as a 'sperm selector' has been suggested from work by Ogasawara et al. (1966) and Lorenz and Ogasawara (1968). The present finding tends to support the postulation that this junction, due to its low level of contractility, may serve as an obstacle for the migration of spermatozoa from the vagina into the upper regions of the genital tract. The walls of the genital tract thicken as they pass the sphincter muscle of the uterovaginal junction to the vagina. Corresponding to the strong musculature, the spontaneous motility of this region exhibited strong but slow contractions (Fig. 6J0). The magnitude of contraction was shown to be highest in the distal middle vagina (1.52 g.) and declined as it aproached the distal vagina (0.21 g.). The finding was in good agreement with that of Sykes (1955) who stated that the motility of the vagina differed from that of the uterus. The only similarity between these two regions may be that the magnitude of contraction of the muscles at the middle portions of uterus and vagina was found to be strongest. This characteristic pattern may be favourable during the transport of an egg from the uterine cavity through the vagina. In spite of the variations in the frequency and magnitude of contractions exhibited by the circular muscles from various regions of the genital tract, it was possible to express their motility in terms of an

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The gradual change in motility pattern can be further noted in the isthmal region where the contraction pattern of the proximal region (Fig 5N) resembles that of the magnal-isthmal junction and the distal region (Fig. 50) more than that of the isthmal-utero junction (Fig. 5P). This change may be attributed to the fact that there are no definite boundaries or striking anatomical differences existing between these two regions (Romanoff and Romanoff, 1949). Frequency of contractions decreased markedly in the isthmal-utero junction. The magnitude, however, remained essentially the same as that of the distal isthmus (Table 2). There were only slight variations in the frequency of contractions (10.5-17.0/min.) found in different regions of the uterus (Fig. 6A-G and Table 2). However, the magnitude of contraction was found to be highest in the proximal region of the middle uterus (Fig. 6C) and lowest in the upper (Fig. 6A, B) and lower (Fig. 6D, E, F, G) uterus. This difference in the magnitude of contraction may be of significance during the transport of an egg from the upper regions of the genital tract into the uterus. The weaker contractions in the upper regions of the uterus may facilitate the easy entrance of an egg into the uterus. Following shell deposition, the egg is forced through the utero-vaginal junction into the vaginal canal with the aid of the longitudinal muscles and by strong contractions of the circular muscles of the middle uterus. The boundary between the uterus and vagina is formed by a strong sphincter muscle (Romanoff and Romanoff, 1969). Motility was extremely weak and irregular (Fig. 6H) at this region. The magnitude of contraction of the proximal utero-vaginal junction was only 0.11 g. and thus the lowest of the entire tract. The magnitude was increased (0.20 g.), however, at the distal region of utero-vaginal junction (Fig. 61).

647

648

T. W. CHEN AND R. O. HAWES

SUMMARY

In vitro motility (frequency and magnitude of contractions) of the genital tract was studied in female Rhode Island Reds. Spontaneous motility of longitudinal muscles from the magnum and uterus was measured using a Grass Model 7 polygraph with birds at 4, 8, 12, 16 20 and 24 weeks of age as well as at 4, 8, and 16 weeks following estrogen priming (diethylstilbestrol dipropionate-4 mg./day for 6 days) of the intact bird. The motility of circular muscles from all regions of the tract of sexually mature females was also measured. The increase of ovary and tract weight with age was positively correlated with the magnitude but not with the frequency of contractions. Frequency of contractions reached a peak between 8 and 16 weeks of age while magnitude of contractions had not peaked at 24 weeks of age. Tract motility in immature females was augmented by exogenous estrogen to resemble that of mature ones. Spontaneous patterns from circular muscles were generally characteristic for each tract region. In this regard the various areas of the uterus exhibited the most variation. An activity gradient was observed,

being greatest in the fimbrial region and lowest in the vaginal region. Activity of the utero-vaginal junction was extremely weak and irregular which may support postulations by other workers that this region acts as a barrier to sperm transport. ACKNOWLEDGMENTS

The authors express their thanks to the Canada Department of Agriculture for grants which made this work possible. REFERENCES Allen, T. E., and L. W. Bobr, 1955. The fertility of fowl spermatozoa in glycerol diluents after intrauterine insemination. Poultry Sci. 34: 1167-1169. Allen, T. E., and G. W. Grigg, 1957. Sperm transport in the fowl. Australian J. Agr. Res. 8: 788-799. Anastassiadis, P. W., W. A. Maw and R. H. Common, 1955. Studies on the glycoproteins of the domestic fowl. 2. The hexosamine content of certain tissues of the sexually immature pullet and some effects thereon of gonadal hormones. Can. J. Biochem. Physiol. 3 3 : 627-637. Brant, J. W. A., and A. V. Nalbandov, 1956. Role of sex hormones in albumen secretion by the oviduct of chickens. Poultry Sci. 35: 692700. Brown, W. O., and N. Jackson, 1960. A study of the composition of the normal and hormonestimulated oviduct of the common fowl, with special reference to its amino acid and nucleic acid content. Poultry Sci. 39:602-611. Chen, T. W., 1964 Effects of a synthetic oxytocin on laboratory and farm animals with special reference to the genital tract. M. Sc. Thesis. McGill University, Montreal. Csapo, A., 1955. The mechanism of myometrial function and its disorder. I n : Modern Trends in Obstetrics and Gynecology, edited by K. Bowes. Butterworth and Co., London. Csapo, A., 1961. Effects of oxytocin substance on the excitability of the uterus. In Oxytocin, Caldeyio-Barcia, R., and H. Heller ed., Oxford. Pergamon Press. 100-121. Csapo, A., and G. W. Corner, 1953. The effect of estrogen on the isometric tension of rabbit uterine strips. Science, 117 : 162-164. Hafez, E.S.E., 1968. Reproduction in Farm Animals 2nd ed. Lea and Febiger, Philadelphia: 235254.

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index (Table 2). With a few exceptions, the indexes calculated were generally greatest in the anterior parts of the tract (e.g. fimbria and infundibulum) and decreased gradually when moving posterially. Index of activity was lowest in the distal lower vaginal region. The observation of such an activity gradient in the genital tract may help explain the transport of ova, or eggs, down the genital tract. It does not however go very far in explaining sperm movement from the storage glands to the infundibulum. Additional work is needed to quantitate blood hormone levels at various stages of egg formation and relate these to oviduct motility.

GENITAL TRACT MOTILITY

ity and embryo normality with site of experimental insemination. J. Reprod: Fert. 16:445455. Mueller, G. C , 1957. A discussion of the mechanism of action of steroid hormones. Cancer Res. 17:490-506. Ogasawara, F. X., F. W. Lorenz and L. W. Bobr, 1966. Distribution of spermatozoa in the oviduct and fertility in domestic birds. III. Intrauterine insemination of semen from low-fecundity cocks. J. Reprod. Fert. 11:33-41. Polge, C , 1951. Functional survival of fowl spermatozoa after freezing at — 79 °C. Nature (London), 167: 949-950. Romanoff, A. L., and A. J. Romanoff, 1949. The Avian Egg. John Wiley and Sons, Inc., New York. Surface, F. M., 1912. Histology of the oviduct of the domestic hen. Maine Agr. Exp. Sta. Bull., 206 :395-430. Sykes, A. H., 1955. The effect of adrenaline on oviduct motility and egg production in the fowl. Poultry Sci. 34: 622-628. Van Tienhoven, A., 1968. Reproductive Physiology of Vertebrates. W. B. Saunders Co., Philadelphia, p. 299-302.

Effects of Early Choline Deficiency on Brain Growth in Chicks JOSEPH M. JOSE, 1 E L I SEIFTER, 2 ' 3 GIUSEPPE RETTURA,3 ALDO CEFALONI3 AND STANLEY M. LEVENSON3 The Department of Surgery, Albert Einstein College of Medicine, Bronx, New York 10461 (Received for publication November 5, 1969)

INTRODUCTION

C

HOLINE deficiency has been studied in laying hens (Nesheim et al., 1967; Burns and Ackerman, 1955) where it has been established that the deficiency leads 1

This paper, based upon data from the thesis of the senior author, is to be presented to the Graduate School of Arts and Sciences of Fordham University, Bronx, New York, in partial fulfillment of the requirements for the Ph.D. degree in Chemistry. 2 Department of Biochemistry, Albert Einstein College of Medicine. 'Department of Surgery, Albert Einstein College of Medicine.

to a decrease in the size of the egg produced, a diminished egg production, and a decreased viability of the fertile egg. The latter phenomenon has led to the characterization of choline as a "hatchability factor" (Balloun, 1956). Although choline deficiency has been the subject of several studies in growing chicks (Fritz et al., 1967; Deeb and Thornton, 1959), the deficient state in the newly hatched bird has not been so extensively studied (Schaeffer et al., 1950). These workers (Schaeffer et al., 1950) observed that a high mortality rate exists for such chicks and that vitamin B 12

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Hafez, E.S.E., 1955. Differential growth of organs and edible meat in the domestic fowl. Poultry Sci. 34:745-753. Farner, D., 1962. Untersuchungen uber die Wirkung von Pharmaka auf Tiere verschiedenen Altero. VI. Die Wirkung von Oxytocin (Syntocinon) und Isoleucyls-Oxytocin (OT198) auf den isolierten Ratten-uterus. Gerontologia, 6: 169-174. Juhn, M., and R. G. Gustavson, 1930. The production of female genital subsidiary characters and plumage sex characters by injection of human placental hormone in fowls. J. Exptl. Zoo. 56:31-61. Kar, A.B., 1947. Responses of the oviduct of immature female fowl to injection of diethylstilbestrol and the mechanism of perforation of the oviduct in the domestic fowl. Poultry Sci. 26: 352-363. Kornfeld, W., and A. V. Nalbandov, 1954. Endocrine influences on the development of the rudimentary gonad of fowl. Endocrinology, 55: 751-761. Lorenz, F. W., and F. X. Ogasawara, 1968. Distribution of spermatozoa in the oviduct and fertility in domestic birds. VI. The relations of fertil-

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