242 



HYPOPHYSIS AND GONADOTROPHIC HORMONES 



the pituitary fractions equated with separate 

 gonadotrophins have been regarded as 

 chemical entities on criteria of purity which 

 have been undergoing revision as advances 

 are made in protein chemistry. 



Several attempts have been made to find 

 extractive procedures b}^ which it would be 

 possible to obtain all or nearly all the pi- 

 tuitary trophic hormones, including the gon- 

 adotrophins, from a common batch of start- 

 ing material (Fevold, 1943; Schwenk, 

 Fleischer and Tolksdorf, 1943; Koenig and 

 King, 1950). Although success in this under- 

 taking would, of course, greatly enhance 

 the potential supply of anterior hypophyseal 

 hormones, such procedures have thus far 

 not proved feasible primarily because of the 

 great differences in the solubility character- 

 istics of the several hormones. Ellis (1958), 

 however, has been able to obtain follicle- 

 stimulating, luteinizing, and thyroid-stimu- 

 lating hormone (TSH) in good yield from 

 common batches of frozen whole sheep pi- 

 tuitaries. 



Studies have been made of the biologic 

 activity of cell-particulate fractions ob- 

 tained by ultracentrifugation of homoge- 

 nized anterior hypophyseal tissue, but it 

 has not been possible to relate gonad-stimu- 

 lating activity to any specific cellular or- 

 ganelle (McShan and Meyer, 1952; Brown 

 and Hess, 1957). 



By the use of histochemical procedures, 

 not in themselves definitive, the secretion of 

 FSH and LH has been ascribed, respec- 

 tively, to specific cell types in the anterior 

 pituitary (see chapter by Purves). 



A. FOLLICLE-STIMULATING HORMONE 



1. Chemical Features 



Chow, van Dyke, Clreep, Rothen and 

 Shedlovsky (1942), working with swine pi- 

 tuitary, obtained an FSH i)rei)aration that 

 was free of other contaminating activities 

 but was physiochemically heterogeneous. In 

 1949 Li, Simpson and Evans ol)tained an 

 FSH fraction (ovine) that was free of othei- 

 trophic factors and that behaved as a single 

 protein by electrophoretic procedures and 

 ultracentrifugation; it was not tested for 

 constant solubility. The hormone FSH be- 

 haves as a protein and there is both chemi- 

 cal and histochemical evidence indicating 



that it is a glycoprotein. FSH from swine 

 and sheep has been variously estimated on 

 the basis of incomplete data to have an iso- 

 electric point of 4.5 and a molecular weight 

 of 70,000 (Li, 1949). 



That the carbohydrate residue in FSH 

 may play an important role in physiologic 

 activity is suggested by the finding that the 

 FSH activity of anterior pituitary extracts 

 is destroyed by salivary amylase and Taka- 

 Diastase (McShan and Meyer, 1938; 

 Abramovitz and Hisaw, 1939). The resist- 

 ance of FSH to proteolysis, however, is nota- 

 ble. In fact, McShan and Meyer (1938), 

 Chen and van Dyke (1939), Chow, Greep 

 and van Dyke (1939), and McShan and 

 Meyer (1940) obtained considerable puri- 

 fication of FSH by means of a selective in- 

 activation or destruction of LH with crude 

 trypsin. More recently, Steelman, Lamont, 

 Dittman and Hawrylewicz (1953) and 

 Steelman, Lamont and Baltes (1955, 1956) 

 have used pancreatin digestion to prepare 

 swine FSH having an activity of 2.5 to 9 

 times the Armour Standard (264-151X). 



In 1950 van Dyke, P'an and Shedlovsky 

 advanced the purification of swine FSH to 

 about 80 to 85 per cent "pure." Since then, 

 Li and Pederson (1952), Steelman and his 

 associates (1953, 1955, 1956), and Leonora, 

 McShan and ]\Ieyer (1956) have made addi- 

 tional improvements in the extraction, re- 

 covery, and purification of FSH with in- 

 creased specific activity. 



Steelman, Kelly, Segaloff and Weber 

 (1956) described the preparation of an "ap- 

 l)arently homogenous follicle-stimulating 

 hormone" with 30 to 50 times the activity 

 of Armour Standard. By placing highly 

 purified FSH on diethylaminoethyl cellulose 

 and utilizing gradient elution, they obtained 

 a 4- to 5-fold concentration of activity as 

 dctcrniined by the augmentation test of 

 Steelman and Pohley (1953). Preliminary 

 study of this FSH in the ultracentrifuge and 

 l)y ])aper electrophoresis revealed no evi- 

 dence of heterogeneity. The molecular 

 weight was calculated to be 29,000 as op- 

 ])Osed to previous estimates. Total carbo- 

 liydrate content was 7 to 8 per cent and was 

 comprised of 50 per cent hexosamine and 

 detectable quantities of mannose, galactose, 

 and fucosc. Tests for LH, TSH, adrenocorti- 

 cotropliic hormone (ACTH), and somato- 



