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2011, Vol. 6 No. 2, Article 94

 

Distribution of Lipid, Protein and Cholesterol in
Adrenal Gland during Postnatal Development in Buffalo

V. Kumar, R. S. Sethi* and O. Singh

 

Department of Anatomy and Histology,
GADVASU, Ludhiana, Punjab- 141004, India

 

*Corresponding Author; e-mail address: sethi116@gmail.com

 


ABSTRACT

The study was conducted on the adrenal gland of 29 buffalo calves for one year to investigate the distribution of lipids and proteins. Based on their ages calves were divided into three groups, Group-I (day old to 3 months), Group-II (more than 3 months to 6 months) and Group-III (more than 6 months to 1 year).
A positive reaction to cholesterol was observed in the cortical cells of the gland in Group – I. The concentration of protein was more in the cells of adrenal cortex and medulla in Group – III. The cells of the zona glomerulosa showed higher content of proteins than the cells of zona fasciculata, zona reticularis and foetal cortex in Group – I . However, the concentration of protein was same in all the cortical zones in Group – II and Group – III
The capsule of the gland in all age groups showed a negative to low amount of fine lipid droplets. Sudan black-B positive lipid droplets content was negative to low in the capsule, low in the zona glomerulosa, high in the zona fasciculata and zona reticularis and negative in the outer and inner zones of medulla and in the area separating cortex and medulla which corresponded to the region of degenerating foetal cortex. Increased amount of lipid in the zona fasciculata may be related to secretion of glucocorticoids. The phospholipid content was more in the cortex as compared to the medulla; however, the cholesterol content was moderate in all the cortical zones and negative to low in the medullary cells.

KEY WORDS

Adrenal, buffalo, lipid, protein, cholesterol.

INTRODUCTION

The distribution of lipids in the adrenal gland has been studied previously in various animals (Cater and Lever, 1954), camel (Nagpal et al, 1991), goat fetus (Hakeem et al., 1993), buffalo calves (Singh et al, 1999). The distribution of protein in the adrenal gland of buffalo foetus was investigated by Sethi (2005). Scanty information available on the distribution of lipids and proteins in the adrenal gland of the buffalo calves made the basis for under taking the present study.

MATERIALS AND METHODS

29 buffaloes calves, based on their age were divided into three groups, Group-I (day old to 3 months), Group-II (more than 3 months to 6 months) and Group-III (more than 6 months to 1 year).
Study of Lipid distribution
Fresh unfixed samples of adrenal glands collected from animals were quickly frozen at –20oC in the deep freezer, subjected to cryostat sectioning to obtain 10-12 mm thick sections. These cryostat sections were stained with Sudan Black-B, Baker’s Acid Haematin and Schultz’s method for demonstration of lipids, phospholipids and cholesterol, respectively (Chayen et al 1969).
Study of protein distribution
The glands were fixed in 10 percent neutral buffered formalin. The paraffin blocks were prepared by acetone benzene schedule (Luna, 1968) and sections of 5-6 mm thickness were obtained on glass slide with the help of rotatory microtome. The sections were stained with Haematoxylin and eosin (Luna, loc. cit).

RESULTS AND DISCUSSION

Total lipid and phospholipid
The capsule of the gland in all age groups showed a negative to low amount of fine lipid droplets (Fig. 1) Hakeem et al (1993b) reported similar findings in goat. The low amounts of sudanophilic lipid droplets in the cells of zona glomerulosa and high amount of lipid droplets in the cells of zona fasciculata observed during present study (Figs. 1, 2). Corroborated the earlier reports of Singh et al (1999) in buffalo calves , Nagpal et al (1991) in camel and Bank (1993). in various domestic animals. Increased amount of lipid in the zona fasciculata may be related to secretion of glucocorticoids .Willims (1970)
In the present investigation, Sudanophilic lipid droplets were found to be absent in the outer and inner medullary zone in the adrenal gland of one month old buffalo calves (Fig. 1) .these findings were in agreement with earlier reports of Singh et al (1999) in buffalo calves and Hakeem et al (1993b) in goat. However, Keene and Hewer (1927) reported low amount of lipid droplets in certain cells of human adrenal medulla. The area separating the cortex and medulla did not show any lipid content which corresponded to the region of the degenerating remains of the foetal cortex (Fig. 2) . (Keene and Hewer 1927). reported similar findings in the human adrenal gland
Phospholipid content was more in the cortex as compared to the medulla (Fig. 3).
Cholesterol
A positive reaction to cholesterol was observed in the cortical cells of gland in Group – I. The moderate amount of cholesterol was observed in the cortex; however, the reaction was low to negligible in the cells of adrenal medulla (Fig. 4). Sethi (2005) reported low to moderate reaction in the definite cortex of buffalo foetus. Where as Greep (1966) observed high amount of cholesterol in the cells of zona fasciculata in the adrenal gland of cow and related it to the steroid secretion activity of cortical cells. The presence of lipid in zona fasciculata may be related to secretion of glucocorticoids (Willims 1970).
Basic protein
The Bromphenol blue positive reaction was observed throughout the adrenal gland indicating the presence of basic protein. The concentration of protein was more in the cells of adrenal cortex and medulla in Group – III suggesting the better differentiation and physiological maturity of the cortico-medullary cells in this age group. The high amount of protein content was also observed in the subcapsular region of the gland. The cells of the zona glomerulosa showed higher content of proteins than the cells of zona fasciculata, zona reticularis and foetal cortex in Group – I (Fig. 5). However, the concentration of protein was same in all the cortical zones in Group – II and Group – III (Figs. 6, 7). The cortical cells contained high concentration of protein than the medullary cells in all the Groups (Fig. 8) These observations were in line with the earlier reports of Sethi( 2005) in buffalo foetu and Vyas and Jacob (1976) in birds .

REFERENCES

  1. Bank WJ. Applied Veterinary Histology, 3rd Edition. Williams and Wilkins, Baltimore 1993, pp. 416-23.

  2. Chayen J, Bitensky L, Butcher RG, Poulter LW. A Guide to Practical Histochemistry. Oliver and Boyd, Edinburg, England 1969, pp. 83-174

  3. Cater DB, Lever JD. The zona intermedia of the adrenal cortex. A correlation of possible functional significance with development, morphology and histochemistry. Journal of Anatomy 1954; 88: 437-454.

  4. Greep OR. Histology, 2nd edition. McGraw Hill Book, New York. 1966, pp. 798-812

  5. Hakeem NA, Sulochna S, Sharma GP, Murthy CT. Histo-chemical studies on the adrenal gland of the common Indian goats (Capra hircus). Indian journal of Animal health 1993b; 32(2): 101-103.

  6. Keene MFL, Hewer EE. Observations on the development of the human Suprarenal gland. Journal of Anatomy 1927; 61: 302-324.

  7. Luna LG. Manual of Histological Staining Methods of the Armed Forces Institute of Pathology. 3rd Edition. McGraw Hill Book Company, New York, USA 1968, pp. 34-157

  8. Nagpal SK, Sudhakar LS, Dhingra LD, Singh Y, Histomorphology of adrenal cortex of camel. Indian Journal of Animal Sciences 1991; 61 (2): 172-175.

  9. Sethi RS. Study on prenatal development of adrenal gland in buffalo. Ph.D. Dissertation submitted to Punjab Agricultural University, Ludhiana, India. 2005.

  10. Singh O, Malhi PS, Singh J. Distribution of enzymes and lipids in the adrenal gland of the buffalo (Bubalus bubalis). Buffalo Journal 1999; 3: 385-389.

  11. Vyas DK, Jacob D. Seasonal study of the adrenal gland of some Indian avian species. Acta Anatomica 1976; 95: 518-528. Willims RH. Textbook of endocrinology. 4th Edition W B Saunders Company, Philadelphia. 1970, pp. 287-290.


FIGURES

Plate 1

Distribution of Lipid, Protein and Cholesterol in

Fig 1-4. Adrenal gland showing 1. Lipid content in the capsule (C), zona glomerulosa (G), zona fasciculata (F) and foetal cortex (f) and no lipid contents are seen in the medulla (M) in one month old buffalo calf Sudan Black B stain x 40. 2. Lipid contents in zona glomerulosa (G) and zona fasciculata (F) in higher magnification. Sudan Blank B stains x 100. 3. phospholipid content in zona glomerulosa (G), zona fasciculata (F) and medulla (M) one month old buffalo calf. Baker’s acid Haematin stain x 40. 4. Moderate amount of cholesterol in cortex and low to negligible in the medulla (M) in one month old buffalo calf. Schultz’s stain x 80.

 

Plate 2

Distribution of Lipid, Protein and Cholesterol in

Fig 5-8. Adrenal gland showing 5. concentration of protein in the capsule (C), zona glomerulosa (G), foetal cortex (f) and medulla (M) in day old buffalo calf. Bromphenol blue stain x 100. 6. concentration of protein in capsule (C), zona granulose (G), zona fasciculata (F) and foetal cortex (f) in four months old buffalo calf. Bromphenol blue stain x 100. 7. concentration of proteins in capsule (C), zona glomerulosa (G), zona fasciculata (F) and zona reticularis (R) in nine month old buffalo. Bromphenol blue stain x 100. 8. concentration of protein in the cells of zona reticularis (R) than medullary cells (M) in four months old buffalo calf. Bromphenol blue stain x 100.

 

 


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