Anat Cell Biol.  2018 Mar;51(1):52-61. 10.5115/acb.2018.51.1.52.

Ameliorative effect of vitamin B12 on seminiferous epithelium of cimetidine-treated rats: a histopathological, immunohistochemical and ultrastructural study

Affiliations
  • 1Department of Anatomy and Embryology, Faculty of Medicine, Mansoura University, Mansoura, Egypt. meladl@sharjah.ac.ae
  • 2Department of Anatomy and Embryology, Faculty of Medicine, Taibah University, Madinah, Saudi Arabia.
  • 3Department of Basic Medical Sciences, College of Medicine, University of Sharjah, Sharjah, UAE.

Abstract

Cimetidine is an H2 receptor antagonist that has an antiandrogenic effect. It intervenes with the conversion of testosterone into estrogen in the Sertoli cells with accompanying testicular structural changes. In the present study, the microscopic and the ultrastructural changes induced by cimetidine and the effect of vitamin B12 as a protective agent on rat testes were studied. Immunoexpression of estrogen receptor β (ERβ) in testes was evaluated. Twenty-four adult male rats were divided into four groups: control, cimetidine-treated, vitamin B12 treated, and combined cimetidine and vitamin B12 treated. The experimental rats were administered with cimetidine and/or vitamin B12 for 52 days. Group II rats showed marked atrophy of the seminiferous tubules with a significant increase in tubular diameter and decrease in the tubular luminal and epithelial areas. Ultrastructure of this group showed irregular Sertoli cells with basal cytoplasmic vacuolation and significantly thickened basement membrane. ERβ immunoexpression was similar to controls. Group III rats showed near normal seminiferous tubular structures with minimal cellular alterations and the immunoreactivity of the testicular sections was very close to normal. However, group IV rats showed markedly immunopositive detached cells, spermatids, and primary spermatocytes. Cimetidine interferes with the control of spermatogenesis as evidenced by microscopic and ultrastructural studies and affection of ERβ receptors and vitamin B12 has a protective action against this harmful effect.

Keyword

Cimetidine; Rats; Seminiferous tubules; Ultrastructural; Vitamin B12

MeSH Terms

Adult
Animals
Basement Membrane
Cimetidine
Cytoplasm
Estrogens
Humans
Male
Phenobarbital
Rats*
Seminiferous Epithelium*
Seminiferous Tubules
Sertoli Cells
Spermatids
Spermatocytes
Spermatogenesis
Testis
Testosterone
Vitamin B 12*
Vitamins*
Cimetidine
Estrogens
Phenobarbital
Testosterone
Vitamin B 12
Vitamins

Figure

  • Fig. 1 Seminiferous tubular shortest diameter, total tubular area, area of the seminiferous epithelium and luminal area: series 1 is a seminiferous tubular shortest diameter, series 2 is the total tubular area, series 3 is the area of the seminiferous epithelium, and series 4 is the luminal area

  • Fig. 2 Photomicrographs of sections of adult rat testis stained by hematoxylin and eosin demonstrating. (A) The control group showing normal seminiferous tubules and interstitial tissue and Leydig cells. The tubular germinal epithelium is formed of different types of spermatogenic cells; spermatogonia, primary spermatocytes, spermatids and sperms (×100). L, Leydig cells; SG, spermatogonia; SC, primary spermatocytes; ST, spermatids, sperms; Sz, spermatids. (B) The control group showing the normal tubular germinal epithelium formed of different types of spermatogenic cells: spermatogonia, primary spermatocytes, and spermatids, sperms (×400). (C) The cimetidine-treated group showing the seminiferous tubular epithelium (spermatogonia, primary spermatocytes, and spermatids) and interstitial tissue and Leydig cells. Marked atrophy of the seminiferous tubules with epithelial disorganization and reduced thickness of germinal epithelium is noted (×100). (D) Higher magnification of panel (C), the seminiferous tubules are damaged with darkly stained pyknotic nuclei (arrowheads). Many vacuoles of variable size are noticed in the germinal epithelium (arrows). Irregular basement membrane with wide interstitial spaces containing (×400). (E) The cimetidine and vitamin B12–treated group showing the seminiferous tubules, the interstitial tissue and Leydig cells. The germinal epithelium (spermatogonia, primary spermatocytes, and spermatids, sperms) is organized in concentric layers (×100). Scale bars=50 µm (A, C, E), 25 µm (B, D).

  • Fig. 3 Photomicrographs of sections stained with immunohistochemistry for detection of estrogen receptor β demonstrating. (A) The uterus (positive control) showing strong immunolabeling in the epithelium and endometrial glands (arrowheads). The connective and muscle tissue showed weak and moderate immunopositivity, respectively (×100). (B) Higher magnification of panel (A) (×400). (C) The testis of control rat showing evident cytoplasmic immunostaining for estrogen receptors in all layers of the germinal epithelium (arrowheads) including spermatogonia, primary spermatocytes, and spermatids (×400). (D) The testis of cimetidine-treated rat showing strong immunopositivity in both nuclei and cytoplasm of spermatids (arrowheads), in the cytoplasm of primary spermatocytes and in the germ cell of the basal layer (×400). P, primary spermatocytes; S, basal layer. (E) The testis of cimetidine-treated rat showing strong immunopositivity in both nuclei and cytoplasm of spermatids (arrowhead), in the cytoplasm of primary spermatocytes and in the germ cell of the basal layer (×400). Scale bars=50 µm (A), 25 µm (B–E).

  • Fig. 4 Photoelectron micrographs of the rat testis demonstrating. (A) A control rat, showing the basement tubular membrane of the seminiferous tubules and myoid cells, primary spermatogonium, Sertoli cell cytoplasm with a large nucleus, homogenous nucleoplasm and prominent nucleolus. Its cytoplasm contains mitochondria. (B) A control rat showing Sertoli cell and spermatogonium with large round nucleus. (C) A treated rat testis with cimetidine, showing Sertoli cell resting on a basement membrane, with flat myoid cells. The cell has folded nuclear membrane with large cytoplasmic vacuole and lysosomes and swollen mitochondria with loss of the homogenicity of the nucleoplasm. (D) A treated rat testis with cimetidine, showing spermatogonia with an irregular nuclear membrane, cytoplasmic vacuoles, distorted mitochondria, and smooth endoplasmic reticulum. Note the fragments of the Sertoli cells in-between the germ cells with dark intercellular fragments and vacuolations (arrow). (E) Group III rat testis, showing Sertoli cell resting on the basement membrane with flat myoid cells. (F) Group IV showing the Sertoli cells resting on the tubular basement membrane with myoid cell. The Sertoli cell nucleus appears homogenous with vacuolated cytoplasm containing mitochondria, tubular endoplasmic reticulum, and lysosomes. (G) Group IV showing primary spermatocytes with a nucleus with clumped chromatin, dilated Golgi apparatus, mitochondria, and tubular endoplasmic reticulum. BM, basement tubular membrane; M, myoid cells; PS, primary spermatogonium; S, Sertoli cell; Mt, mitochondria; SP, spermatogonium; V, cytoplasmic vacuoles; L, lysosomes; ER, endoplasmic reticulum; ch, chromatin; GA, Golgi apparatus. Scale bars=5 µm (A–G).


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