Surks M I, DeFesi C R
Endocrinology. 1977 Sep;101(3):946-58. doi: 10.1210/endo-101-3-946.
A method combining enzymatic cellular dispersal, direct cell counting, differential cell counts at the electron microscope level and DNA determinations was devised and employed for determination of the cell numbers of each anterior pituitary cell type in euthyroid (E) and hypothyroid Tx) rats. Pituitaries from Tx rats had increased cell number as demonstrated by a mean 33.7% increase in DNA content (microgram DNA/pituitary). Total cells increased from (3.14 +/- 0.36) X 10(6) in E rats to (3.98 +/- 0.27) X 10(6) in Tx rats. P less than 0.005. The cellular DNA content (g/cell) in E rats ,10.84 +/- 0.63 (SD), was indistinguishable statistically from that of Tx rats, 11.24 +/- 0.52. Cell distribution among various pituitary cell types was virtually identical when determined in pellets from dispersed cells and randomized solid tissue from the same groups of E and Tx rats. These data indicated that there was no selective cell loss during the cell dispersion procedure. Major changes in Tx rats compared to E rats were a marked increase in percentage of thyrotrophs, from 10.7 +/- 1.75 (E) to 34.4 +/- 1.0 (Tx), and a decrease in percentage of somatotrophs, from 55.3 +/- 1.82 to 15.3 +/- 0.97. The calculated cell distribution showed that the number of thyrotrophs increased from 0.34 +/- 0.02 to 1.37 +/- 0.05 millions per pituitary and somatotrophs decreased from 1.74 +/- 0.11 to 0.61 +/- 0.02 millions in hypothyroid rats. The method described herein thus provides a quantitative estimate of changes in pituitary cell populations in different hormonal states and should be useful in studies of the kinetics of pituitary cell replication and removal.
设计并采用了一种结合酶促细胞分散、直接细胞计数、电子显微镜水平的差异细胞计数和DNA测定的方法,用于测定甲状腺功能正常(E)和甲状腺功能减退(Tx)大鼠中每种垂体前叶细胞类型的细胞数量。Tx大鼠的垂体细胞数量增加,DNA含量(微克DNA/垂体)平均增加33.7%证明了这一点。总细胞数从E大鼠的(3.14±0.36)×10⁶增加到Tx大鼠的(3.98±0.27)×10⁶。P<0.005。E大鼠的细胞DNA含量(克/细胞)为10.84±0.63(标准差),与Tx大鼠的11.24±0.52在统计学上无差异。当从相同组的E和Tx大鼠的分散细胞沉淀和随机固体组织中测定时,各种垂体细胞类型之间的细胞分布几乎相同。这些数据表明,在细胞分散过程中没有选择性细胞损失。与E大鼠相比,Tx大鼠的主要变化是促甲状腺激素细胞百分比显著增加,从10.7±1.75(E)增加到34.4±1.0(Tx),生长激素细胞百分比降低,从55.3±1.82降低到15.3±0.97。计算出的细胞分布表明,甲状腺功能减退大鼠中促甲状腺激素细胞的数量从每个垂体0.34±0.02增加到1.37±0.05百万,生长激素细胞从1.74±0.11减少到0.61±0.02百万。因此,本文所述方法提供了不同激素状态下垂体细胞群体变化的定量估计,应有助于垂体细胞复制和清除动力学的研究。