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ENGLISH SUMMARY .
Subject of the thesis :
Effect of Haloperidol ( Hadol - decanoso ) antidepressant drug on the development of some organs in chick embryo.
Introduction and review of Literature .
We enumerated the references specially of depressant ; from identification its and classification of antidepressant and antineuroleptic drugs .The effect of depresstion on ladies on deferent age , for special pregnant women's and on the post-partum period . The general effect of antidepressant on animals and on some region and function of the body . The absorption and fat of the drug . The references of haloperidol drug ; (characterization ; mechanization ; absorption ; distribution ; method of the expulsion ; side effects and some reference about the effect of the drug on chick , mouse , rats , rabbit, dogs and human generally and on chosen organs specially . Next enumeration of the reference of normal development of retina , kidney and liver of chick embryo specially , and on birds , animals , mammals and human generally .
Material and methods .
First : material used in this investigation .
(780 egg) were fertile used in this (work), and were divided into two groups :
A) : first group ( control) :
This group were used to study normal development and structure of the retina , kidney and liver , which contain (200 egg) ,and open the eggs and extracted the embryos at ( 5,6,7,8,9 ,10,12,14,16 and 19-days) of incubation .
B) : second group ( treated) :
This group contain (580 egg fertile) and were injecting by equivalent therapeutic dose of haloperidol (0.25 mg/egg) in air sac space by a dose . This group subdivided into 5-division according to time of injection and repeated doses ;
(1)- subdivision –(T0) : injection in zero-day before incubation by one dose (single dose) , and the embryo collected at the same day of the control group .
(2) - subdivision –(T1) : injection in the first week at the 5-day by one dose ,and the embryo collected in same day of control group and (T0) exclude the 5-day to compare between embryos .
(3) - subdivision –(T2) : injection in second week at the 11-day by one dose and the embryo collected at the (12,14,16,19-days).
(4) - subdivision –(T3) : injection in third week at 15-day by one dose and the embryo collected at (16,19-days).
(5) - subdivision –(T4) : (the group of repeated doses ) injection in first , second and third week in 5,11,15-days by one dose and the embryo collected at (12,14, -days) which taken double dose . And collected the embryo of (16,19 –days) which taken triad dose .
second : methods used in this investigation .
- The fertilized chick eggs were used and injection by Sesame oil on the control group and by haloperidol drug on treated groups at the air chamber (air-sac) . The eggs were open and determines ages according to selection study days .
- The morphogenesis were studied as contain malformation ; weight of embryos ; definition of the mortality rate on treated and control group and comparing between them and statistic study were done .
- The histological (cellular structural) were studied for retina, kidney and liver , as all embryo fixated in Bouin's solution or formalin saline 10% ,then making the steps of preparation of study histological structure for selective organs of chicken embryo, by following stains : (Haematoxylin and eosin stain to study general form and cellular structure ; Heidenhins iron hematoxylin to study mitotic figure rate of liver cells ; Holms silver stain to study fibers rata on the retina ). The Morphometric measurement were done by using slides of Haematoxylin and eosin of the kidney of the 19-day by computer system of [Image – pro plus (v 5.1)] . For electron microscope study , fixation liver of at 16-day by gloteraldhyde for making ultrathin sections to study ultra-structure of the liver tissue in all studied groups .
Result.
In this investigation normal development of retina, kidney and liver were studied , then study of the effect of haloperidol drug on morphogenesis , weight , mortality rate and histological structures for selection organs .
First : morphogenesis .
Appeared as a results of this study . Malformation in some embryos such as; Congesting of the body , decreased the length of the body and brain , herniation of the brain (the vault of the skull not formed ) , absent of the eyes or decreased on ambience , absent of the ear vesicles, smaller of the beak, kinking of the neck , deformed the length of lambs , the feather not formed or less appeared and hernia of the gut) . The (T0) subdivision appeared more affected in malformation appearance , then (T1, T4) repeatedly dissecting ; the embryo some morphogenesis changes ; bulging of some organs , congestion of the liver and kidney , where hear colors appeared semi-dark , as a result of internal haemorrhage as comparing with control one .
Second : weight .
After study of this investigation and comparing the weight of treated with control embryo . Decreased of the weight of treated embryo communally . The reduction was significant in (T0) and in (5,6,7 days), also in (T1) the reduction was occure but non-significant in (6,7 –days) , while in (8,9,10,12,14,16, and 19-days) reduction was significant in whole treated groups (T0,T1,T2,T3,T4) . The (T0) group was more affective in loss of weight .Also the decrease of weight was significant in subdivision groups which taking single or repeated dose
Third : mortality rate .
After examine the result of this investigation the mortality rate were higher in (T0) [25%-30%] , then (T4) –[ 15%-35%] , then (T1)-[5%-30%] , while in the (T2 and T3) the ratio ; were less [5%-10%] , and appeared that higher of mortality rate in (T4) which taking repeated doses , than the all subdivisions; excluded the (T0) subgroup which taking the drug before incubation , thus the effect of haloperidol drug depend on times of take dose and repetition of the dose .
Fourth : histological studies .
A) : normal development of chick embryo .
1): normal development of retina.
From this study appearance that the retina appeared in (5-6 days) composed of 3-layers ; pigmented layer , sensory (neurobastic )zone and marginal layer , the development were rapid from posterior part there extended two part anterior gradually . Then the layers differentiated and increased in numbers . In (7-8 –days) the retina appeared differentiated in posterior than the lateral part into layers ; pigmented layer and sensory layers which differentiated to 6-layers : External limiting membrane (E.L.M), Neuroblastic zone (NZ), Inner reticular layer (I.R.L), Ganglion cell layer (G.C.L), Nerve fiber layer (N.F.L) and Internal limiting membrane (I.L.M).[ here the first appearance of ganglion cell which were formed of 4-5 row of cells].In the 8-day the thick of Neuroblastic were fromed of (16-18 row of cells), which differentiated to 3-sublayers ; inner horizontal cells (amacrine cells) and intermediate horizontal cells (bipolar cells) and outer horizontal cells .While the lateral part the retina still formed of 3-layers with increase of their thickness . In the 9-day the numbers of sensory layer were increased to 8-layers as two layers were added (the Hinle,s membrane (H.M) which the characteristic of birds retina and the Layer of visual cells "rods and cones (R&C)". These was the first appearance for them. The development the of layers retina of the lateral and posterior part was differentiated and full development become clearing in the two sides but posterior part was more thick. The inner reticular layer(Neuroblastic previous) increased to (17-19-layers) and the Ganglion cell layer decreased to (3-4-layers) . Continuation of the development in the next ages (10,12,14,16 and 19-days) with observance that the retina increased in thick from 5-day arrival to 10-day of incubation , than increased in thick (especially nuclear layer and ganglion layer where the thick of nuclear layer become (9-10 layers) and ganglion layer become (1-2 layers) in 19-day) , beside the increased of the fibrotic layers; (Inner reticular layer and Nerve fiber layer. As become easily to differentiation the Inner reticular to several parallel rows) tell full development .
2): normal development of kidney.
Mesonephrose .
The light microscope of this investigation revealed that the mesonephrose could be seen by the 5-day ,as an aggregate of renal corpuscles and renal tubules as age advance this components increased in numbers and some units of renal corpuscles had seen , The medial end of renal tubule invaginated by blood vessels to formed the glomerulus as cup-form contain of entangled aggregation from blood capillary , and Bowman's capsule there formed and it which contain two layers outer parietal layer from of simple squams cells and inner visceral layer from of rounded cells adherent to blood vessels , between them appeared the capsular space (parenthesis-form) , the renal corpuscles extended in between the renal corpuscles with numerous sizes and forms and connective tissue and mesen-chymal cells between them . The mesonephrose continued its development at (7,8,9-days) by increased the size and proliferation of its the units and more differentiation of its cells . At 7-day to the renal tubules differented into 3 segment ; proximal and distal segments in between them the transitional one . Then more differentiation occur to proximal , distal tubules and loop of Henle's in 8-day . From 10-day the renal units appeared less in numbers than previous age, as large cavities appeared between this tubules and the lumen of renal tubules more dilatation than previous age, the glomerulus occupy the medial side region the mesonephrose , while the tubules occupy most of lateral part area of mesonephrose . Age advance (16-19-day) reduce the mesonephrose size and become as a remnant from renal corpuscles and tubules.
The mesonephric duct had been appeared at the 6-day as a rounded duct on dorsolateral side of the mesonephrose which formed of columnar cell rested on clear basement membrane, and surroundings by rounded cells and mesenchymal spindle-form cells, this development continued as age advanced till to 10-day , and as the degeneration of the mesonephrose occur less appearance and the duct could not be seen .
The sign of degeneration of mesonephrose from (9-10-days) occurred as a tissue degenerated and necrosis of cell . The cell depress appeared between the renal compounds . The renal tubules and glomerulus, continued the degenerative shapes as age advanced. At (16-19-day) degenerative changes in the whole compounds of mesonephrose become more and clear as the metanephrose is the active and function kidney .
Metanephrose .
The result this study revealed ;that the metanephrose appeared early at 6-day as aggregates of small vesicles and collecting tubules which surrounding by metanephrogenic blastema which differentiation into two zones ; inner nephrogenic zone as inner crowded cells and outer nephrogenic zone less crowded cells . The primordium of the glomerulei appeared at (9-10-days) and in the 10-day the lobeulated of the metanephrose begin as differentiation of some of this compounds. The central vein , ureter and interlobular veins, at the 12-14-day had been differentiation , also the renal corpuscles of metanephrose . The differentiation of the kidneys into outer cortex and inner medulla occurred . The kidney appeared as a long douple superiorly organs surrounding by thin fibrotic capsule ,and elongate as from lungs to loccye inferiorly in embryo . From 16-19-day the kidney could be seen by naked eye , as a lubulated organs .It had ( anterior ,medial and posterior lube) each lube is pear shaped and formed of lobules which separated by fibrotic septa and contain a blood vessels and ureter each lube from of two parts lobular cortex and inner medulla , the kidney contain numerous of renal compounds , where the renal tubules differentiation into ; proximal, distal , loop of Henle's and collecting tubules .The renal corpuscle differentiation into two type of glomerulus (an outer cortical glomeruli in the cortex and an inner medullary glomerulei) which surrounded by Bowman's capsule which differentiated into two layers; outer parietal layer and inner visceral layer in between them capsular space . All the components of the metanephrose ; ureter, central veins, interlobular veins completed differentiation . At the 19-day as the cortical part contain the renal corpuscle , convoluted tubules ;proximal and distal, while the tubules of medulary part appeared less numbers and contain the collecting tubules and loop of Henle's. The ureter had seen which appeared at the 14-day of incubation veiled by transitional epithelial ,surrounding as lined by smooth muscle which divided into inner longitudinal layer and outer circular layer , surrounding by fibrotic connective tissues and mesenchymal cells .
2): normal development of liver .
By light microscope .
The result of this investigation revealed that ; the liver appeared in the 5-day of incubation , t formed of two lobes large right lobe and small left lobe connected by wide (empty space) intermediate portion . And the liver surrounded by fibrotic connective tissue capsule , the liver appeared histological contain aggregate from of cellular cords which differentiation into solid cords ( formed of two rows of polygonal cells) and lumenated cords (fromed of one row of pyramidal cells (8-10) cells around small lumen)in between them wide tortuous blood sinusoids which lined by two type of cells ; flat endothelial cells and kupffer cells . As age advanced 7-8-day of incubation liver cells (hepatocytes) increased as compared to blood spaces , that main increased in the cell density and narrowing of blood sinusoids . As more development of hepatic cords , the intermediate portion begin to grow and full of cells , with predominated of the lumenated cords than the solid cords which formed of (6-8 cells) .The hepatic cords appeared more crowded at the central region of the two lobes of the liver than the peripheral parts. As age advanced from 9-14-day the liver increased in size and increased the numbers of solid and lumenated cords with predominated lumenated cords than the solid one . The lumenated cords decrease the its density to (5-6 cells) about each lumen . At 16-day the size of blood sinusoids decreased gradually . And the mitotic figures of hepatocytes decrease . From 16-19- days the hepatic cords become compacted so there are difficult to different the solid or lumenated cords . Several of cellular mitotic figures has been observed in numerous stages of the hepatocytes , the mitotic figures began from the 5-day of incubation and decrees as age advance. At 7-and 9-days the mitotic figures decreased as age advanced .