The present study was conducted on 18 gravid uteri from healthy goats (Capra hircus) of non descript breed procured from local slaughter house. The uteri were divided into three groups according to their gestational ages as Gr I (0-50 days), Gr II (51-100 days) and Gr III (101 till term). Surface epithelium, stratum compactum and stroma of endometrium, circular and longitudinal muscle layers in myometrium, blood vessels in stratum vasculare and perimetrium during early pregnancy show mild activity with the acid and alkaline phosphatase which increased with the advancement of the pregnancy. The endometrial glands showed mild to intense activity of alkaline and acid phosphatase during initial stage of pregnancy. The activity of alkaline phosphatase increased with the advancement of pregnancy. The activity of acid phosphatase was moderate to intense during mid stage and moderate during advance stage of pregnancy.
A large amount of monosacharides are known to be produced in the uterus under the influence of oestrogen (Zachariae, 1958). The acid phosphatase (orthophosphoric-monoester phosphohydrolase, acid optimum) and alkaline phosphatase (orthophosphoric-monoester phosphohydrolase, alkaline optimum) is closely involved in the metabolism of monosacharides (Bachhawat et al., 1972; Valocky et al., 2007). Acid phosphatase (ACP) may be involved in the provision of energy for organelle destruction. Acid phosphatase release has been associated with lysosomal involvement in implantation and placentation in the sheep (Boshier 1969), man (Contractor et al., 1977) and mouse (Moulton et al., 1978). The frequent association of the enzyme alkaline phosphatase with monosaccharides in the tissue was observed (Moog and Wenger, 1952). Alkaline phosphatase may be associated with the transport of nutrients across and the transfer of energy within the microvillar membrane of the absorptive surface (Fernley, 1971). Alkaline phosphatase also involves in the luminal epithelium and trophoblast of the sheep has been associated with carbohydrate metabolism and the production of trophoblastic fibrinoid (Boshier 1969). The decidual cells showed intense alkaline phosphatase activity during pregnancy in rat (Pritchard, 1947).
ALP and ACP are two different enzymes which show considerable alterations depending on the volume of luminal secretion (those secretions which are essential for embryo development) in luteal phase. Alkaline phosphatase (ALP) is known as a major sialoprotein and is mainly localized on the basal and apical cell membrane and cytoplasm of the epithelial and glandular epithelium (Fleming et al., 1995). This enzyme plays very important role in early phases of implantation (Lindhard et al., 2002). Especially in luteal phase, the acid phosphatase (ACP) activities are responsible for hydrolysis of organic phosphoesterase (Leiser and Wille, 1975b).
In the view of above observation the objective of present study was to elucidate the distribution of alkaline phosphatase and acid phosphatase activity in the different parts of the gravid uterus of non-descript breed of the goat.
Materials and Methods
The present study was conducted on the 18 gravid uteri procured from healthy goats (Capra hircus) of non descript breed. An approval from the animal ethic committee DUVASU, Mathura (U.P.) was obtained prior to the commencement of the study. Immediately, after collection the umbilical cord was ligated and foeti were cleaned with the help of cotton soaked with water to remove the amniotic fluid. Each foeti was measured for its crown rump length (CRL) in centimeters with the help of nylon tape as per the technique described by Harvey (1959) and weighed in grams with the aid of electronic weighing balance machine. The approximate age of foeti above 30 days was estimated by using the formula derived by Singh et al. (1979) in goat. Below 30 days approximate age of foeti was estimated by its crown-rump length. The uteri were divided into three groups according to their gestational ages as Gr I (0-50 days), Gr II (51-100 days) and Gr III (101 till term). In the uteri of group-I, II and III a small piece of both uterine horns (apex and base both side) and body. Tissue were collected and fixed in 10% neural buffered formalin. Paraffin blocks were prepared by acetone benzene schedule by using low melting point paraffin of 46-480c and 15 µm thick sections were obtained on the glass slides with the help of rotary microtome. The sections were stained with Gomori’s method for study of alkaline phosphatase and acid phosphatase (Luna 1968).
Results and Discussion
Surface epithelium, stratum compactum and stroma of endometrium, circular and longitudinal muscle layers in myometrium, blood vessels in stratum vasculare and perimetrium during early pregnancy showed mild activity with the alkaline phosphatase (Fig.1; Table 1).
Fig. 1: Photomicrograph showing alkaline phosphatase reaction in the surface epithelium (endometrium) of goat at 33 days of pregnancy (arrow) show mild activity. (Gomori stain, 10X)
Table 1: Histochemical variation of alkaline and acid phosphatase in the gravid horn of uterus during different stage of pregnancy in goat
|Enzymes||Alkaline Phosphatase||Acid Phosphatase|
|Parts of Gravid uterus / Group||Ist||IInd||IIIrd||Ist||IInd||IIIrd|
|Stratum compactum||+||++||± to +||+||+||+|
|Stroma||± to +||± to +||±||+||±||+|
|Endometrial Glands||Neck segment||+ to +++||+ to +++||+++||+ to +++||++ to +++||+|
|Middle segment||+ to +++||+ to +++||++ to +++||+++||++||++|
|Basal segment||++||+++||+++||+++||+ to +++||++|
|Myometrium||Circular muscle layer||± to +||±||±||+||+||+|
|Longitudinal muscle layer||+||± to +||±||+||+||+|
|Blood Vessels in Stratum Vasculare||+||± to +||± to +||+||±||+|
|Perimetrium||± to +||+||+||+||+||++|
(± to +) very mild, (+) mild, (++) moderate, (+++) intense, (-) no reaction
In the surface epithelium and stratum compactum, the alkaline phosphatase activity increased with the advancement of pregnancy (Table 1). Almost similar activities were observed in the stroma of endometrium, circular and longitudinal muscle layer in myometrium, blood vessels in stratum vasculare during all stages of the pregnancy similar observations were made by Chandra (1973) in buffalo. Rawal, (2016) observed the ALK activity in the uterus of rat. Maureen and Rolls (1980) in rat observed that alkaline phosphatase activity was increased during pregnancy.
In the perimetrium very mild activity of the ALK were observed during initial stage of the pregnancy which become mild during mid and advance stages of the pregnancy. The neck and middle segment of the endometrial glands showed moderate to intense activity of ALK which decreased with the advancement of the pregnancy (Table 1). At initial stage of pregnancy, the basal segment of endometrial glands showed moderate activities which become intense during mid and advance stages of pregnancy (Fig.2).
Fig. 2: Photomicrograph showing alkaline phosphatase reaction in the endometrial glands of goat at 120 days of pregnancy (arrow) show moderate to intense activity. (Gomori stain, 10X)
Similar observations were made by Chandra (1973) in pregnant buffaloes. Hafez and White (1968) in ewe mentioned that alkaline phosphatase activity was present in apical cell membrane of the opposing trophoblast and uterine epithelium. Zamiri (1980) in ewe observed that alkaline phosphatase activity in the luminal epithelium, uterine glands and maternal caruncles was high and did not change during pregnancy. Shabrooz et al. (2013) is observed that in river buffalo the more ALK activity in the glandular epithelium than the endometrial epithelium. Surface epithelium, stratum compactum and stroma of endometrium, circular and longitudinal muscle layers of myometrium, blood vessels of stratum vasculare and perimetrium of the gravid uterus showed mild reaction of the acid phosphatase during whole pregnancy (Table 1). In advance stage of pregnancy the surface epithelium of the endometrium show moderate activity of ACP present and similar observations were made by Maureen and Rolls (1980) in rat. However high level of acid phosphatase activity in uterine surface epithelium was reported during early pregnancy by Hafez and White (1968) and Boshier (1969) in ewe. Boshier (1969) further mentioned that this enzyme was responsible for the release of lysosomal hydrolysin.
Fig. 3: Photomicrograph showing acid phosphatase reaction in the surface epithelium (endometrium) of goat at 120 days of pregnancy (arrow) show mild to moderate activity. (Gomori stain, 10X)
In the present study the neck and middle segments of the endometrial glands during initial and mid stages of the pregnancy showed moderate to intense activity. The basal segment showed intense to moderate activity of the acid phosphatase (Table 1). Chandra (1973) in buffalo observed that acid phosphatase showed mild reaction in neck segment and mild to intense reaction in middle and basal segments. Shabrooz et al. (2013) is observed that in river buffalo the more ACP activity in the glandular epithelium than the endometrial epithelium. Bachhawat et al.(1972) reported that acid phosphatase and alkaline phosphatase is closely involved in the metabolism of monosaccarides. Maureen and Rolls (1980) in rat observed that acid phosphatase was increased during pregnancy and suggested that increase in acid and alkaline phosphatase activities probably, reflects the need for increased nutrient absorption.
CONCLUSION: PLEASE ADD CONCLUSION