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Light and Electron Microscopic Observations on Cytological Changes in the Anterior Hypophysis of the Mouse Fol- Lowing Exposure to Cold

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Light and Electron Microscopic Observations on Cytological Changes in the Anterior Hypophysis of the Mouse Fol- Lowing Exposure to Cold Okajimas Fol. anat. jap., 43: 263-289, 1967 Light and Electron Microscopic Observations on Cytological Changes in the Anterior Hypophysis of the Mouse Fol- lowing Exposure to Cold By Wen-Der Wu Department of Anatomy, Nagoya University School of Medicine, Nagoya, Japan (Director : Prof. Dr. K az u mar o Yamada) Introduction The cytological observations on physical and experimental con- ditions in the anterior hypophysis of human and vertebrates were undertaken by many investigators (S c h O n e m a n n 1892, Da w son and Friedgood '38, Romeis '40, Dawson '46, Goldberg and C h a i k o f f '52 and others). During these decades, in this laboratory, studies on relationships between specific cellular structures and a functional significance of secretory cells in the anterior hypophysis have been consecutively made and the considerable results were obtained (Y a m a d a '37a, '37b and '43, Yamada et al., '56, S a n o '56 and others). In the area of electron microscope too, the fine structures of the functional cells of the anterior lobe were observed and the morphological classification of cell types correlate to hormone production has been undertaken these years. Especially, an electron microscopical study on the possible site of the production of ACTH in the anterior pituitary was reported by Yamada and Yamashita in 1966. Since the general adaptation syndrome described by Se 1 ye ('37), many workers have attempted to identify a specific type of cell in the hypophysis. This syndrome is occurred to hyper-produc- tion of ACTH in the anterior hypophysis as a result of non-specific stress. It is known that atrophy of sex organs and hypofunction of thyroid glands in the animals exposed to stresses for long time take place as part of the general adaptation reactions (S e 1 y e, '36, '37, '39, '40, '47). The cytological studies of the anterior pituitary of the non- 263 264Wen-Der Wu specific stressed animals have been made by many workers in order to search a cellular source of ACTH (M a 11 g r e n, '48, Halm i, '50, Finer t y et al. and others). B a i 11 i f ('38) and M c N a r y ('57) reported the cytological changes in the anterior hypophysis of the animals following exposure to cold. In this laboratory, F u j i n o ('62) made an extensive observations of the cytological changes in the anterior pituitary of the mouse follow- ing chronic exposure to heat considered to be a kind of non-specific stress. The present study was undertaken to investigate an influence of cold stress in the anterior pituitary of the mouse with light and electron microscope. Materials and Methods Ninety-four male and one hundred and five female DD strain mice between 60 to 90 days of age were exposed to cold ranging from 3°C to 5°C for periods of 2, 4, 6, 8, 12, 16, 18, 20 and 24 hours and 2, 4, 5, 11, 20, 32, 34, 36, 42, 53, 60, 71, 86, 97, 110, 120, 135, 140 and 191 days. On the other hand, 17 male and 28 female mice between 60 to 90 days of age were kept with a constant temperature of about 23°C as normal control group. The experimental groups were given food and water ad libitum as same as in normal control animals. For light microscopic observations they were sacrificed under chloroform anesthesia. The pituitary obtained was fixed with Zenker- formalin acetic acid solution, embedded in paraffin and sectioned seri- ally in the sagittal plain at four microns. These sections were pre- pared with Han.sen's hematoxylin-eosin stain, Heidenhain's azan stain, periodic acid Shiff reaction and Gomori's aldehyde fuchsin stain. For electron microscopy, the pituitary was fixed with buffered isotonic one per cent osmium tetroxide (OsO4) (P a 1 a d e, '52) and embedded in Epon 812 (L u f t, '61). The sections were cut with a JUM-4 type ultramicrotome, stained with uranyl acetate (W a t s o n, '58) and examined and photographed with Hitachi HU 11A electron microscope. Observations The secreting cells of the anterior pituitary grand may be divided into seven types : the first type acidophile cell corresponds to growth hormone secreting cell (GH), the second type acidophile cell to luteo- trophic hormone secreting cell (LTH), beta basophile cell to thyroid- stimulating hormone secreting cell (TSH), the first type delta baso- Cytological Changes in the Anterior Hypophysis of the Mouse 265 phile cell to follicle stimulating hormone secreting cell (FSH), the second type delta basophile cell to luteinizing hormone secreting cell (LH), the chromophobe cell to peculiar non-granulated cell (PNG) and adenocorticotrophic hormone secreting cell (ACTH). I. Normal Control Group Acidophile Cell : In light microscopy, the acidophiles are more numerously found in the anterior lobe. The cells are round or oval in shape and various in size and divided into small, medium and large acidophiles. Medium sized acidophiles are predominant in number. They are well granulated with acidophilic granules in the cytoplasm. Medium sized and large acidophiles have negative Golgi image, perinuclear halo and cytoplasmic basophilic substance. The nuclei of acidophiles are situated centrally in the cytoplasm of the small sized cells but eccentrically in medium sized and large ones. The nuclei contain fine chromatin particles in small numbers, and one or two conspicuous nucleoli and show a light vesicular cast. Mitotic figures of acidophiles are rarely seen. In electron microscopy, two types of acidophiles are identified. The First Type Acidophile Cell (Gil Secreting Cell) : This cell is most easily identified and numerously found in electron micro- scopy. The cell contains relatively large electron dense granules, the average diameter is about 350 mp. The lamellae of the endo- plasmic reticulum are well developed, the Golgi apparatus is clearly visible and the appearance of the mitochondria is often characteristic. The Second Type Acidophile Cell (LTH Secreting Cell) : This cell is characterized by the presence of oval or irregularly shaped dense granules. The longest dimention of the granules is 150 to 400 mp. The endoplasmic reticulum is aligned with flattened parallel lamellae and the membrane is richly studded with ribosomes. The Golgi apparatus is not well developed. Beta Basophile Cell : The beta basophiles (TSH), in light micro- scopy, are scattered in small numbers through the anterior lobe. They are polygonal and irregular in shape. Their cytoplasm are filled with fine PAS positive granules. The nuclei of beta basophiles are round, oval or elongated in shape but sometimes wrinkled, and usually smaller than those of the delta basophiles. They contain many chromatin granules and show a dark cast. The TSH cells in the electron microscope are polygonal with fine granules. The granules often line up the peripheral region of 266Wen-Der Wu cell membrane and the maximum diameters of the granules vary between 50 and 200 mg. The Golgi apparatus and the endoplasmic reticulum are relatively poorly developed. Delta Basophile Cell : In light microscopic observations, the delta basophiles are also in small numbers. They are found in clusters in the anterior portion of anterior lobe and scattered in the central area. They are oval or polygonal in shape and filled with fine dust-like PAS positive granules in the cytoplasm. The nuclei of delta basophile cells are round or oval in shape, contain a single large acidophilic nucleole and abundant fine chromatin particles. Under the electron microscope, the delta basophile cells are divided into two types, namely the first type • delta basophile cell (FSH) and the second type delta basophile cell (LH). The FSH cells are characteristic and distinguished from the other cells. The secretory granules are relatively electron-transparent and scattered throughout the cytoplasm. The diameters of the granules vary between 150 and 200 mg. The Golgi complex is well developed and the endoplasmic reticulum is poorly developed. The cells are usually adjacent to the blood vessels. The LH cells are relatively small, round or oval in shape. The granules are distinguished by their heterogeneity of size and much more dense than those of the FSH cells. The average diameter of the granules varies between 100 and 200 mg. These usually accumu- late at the periphery of the cells. The endoplasmic reticulum is not prominent and appears as fine scattered tubules and the Golgi com- plex is moderatly developed. Chromophobe Cell, PNG Cell : In light microscopic observations chromophobes are randomly located in small clusters or individually The cells are various in size and smaller than the chromophile cells and the frequency of appearance is low. They contain few granule:. and flocculent material in the cytoplasm. Many of them show ar indistinct cell boundary. The chromatin-network of the nuclei coars. Mitotic figures of chromophobes are occasionally observed. The PNG cells in electron microscopy, the granules are hardll visible, the Golgi complex is relativery obscure and the mitochondria are poorly developed. ACTH Secreting Cell : In light microscopy, the general morpho logy of ACTH cells resembles that of the first type delta basophile cell (FSH), but the whole cell bodies are much larger than those o FSH cells. They are irregular in shape, in small number and oftei found in the anterior portion of the pituitary gland. The cytoplasn Cytological Changes in the Anterior Hypophysis of the Mouse 267 are filled with fine granules and the nuclei contain abundant fine chromatin particles. In electron microscopy, the ACTH cells are provided with a prominent characteristic in morphology. The shape of this cell is somewhat irregular, polyhedral and elongated. The secretory granules are most characterized by their variable appearances with solid granules and haloed granules or vesicular form with a dense core.
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