A retinal substance was disclosed which was histochemically demonstrable by consecutive treatment with a concentrated solution of sodium bisulfite (15% aqueous solution,24 hours,20° C. ) and Schiff's reagent. Distribution of the substance in anuran rod acromeres (outer segments) corresponds to the site of positive tinctorial response in the peracetic acid Schiff staining, with the use of which comparative experimentation was made in order to obtain analytical data for the underlying mechanism of histochemical reaction of the substance.
The substance in question was learned to be favorably preserved by various aqueous fixatives containing either potassium bichromate or mercury chloride or both in combination or in sequence. Bisulfite Schiff reaction of the substance was effectively blocked by preceding bromination or iodination. So far as the pyridine-insoluble substance in the acromeres is concerned, the reaction was out of the blocking effect of acetylation. In test tube, it was observed that oleic acid gave positive bisulfite Schiff staining, while palmitic acid and stearic acid behaved negatively. Oleic acid after bromine treatment lost the responsibility to the staining. On the basis of such observations it is deduced that the present bisulfite Schiff staining of the retina might be due to possible interaction with ethylenic double bonds.
Pretreatment with semicarbazide, anilin chloride or phenylhydrazine blocked the staining, whereas hydroxylamine was without signifi- cant effect. Coloration resulting fromthe staining reaction is less re- sistant to acid. When bisulfite treatment preceded,, silver reaction became negative, but the affinity to thiazine dyes such as methylene blue and thionin was left unimpaired even under low pH condition. Thionin was responsible for metachromatic basophilia at pH 1.2. Such basophilia was lost by methylation (1 hour, ° C.; 6 hours, ° C. ), and the loss of basophilia was correlated with negative Schiff reaction. Both stainings were similarly affected by 2 days' iodination. Exposure of the substance to the air at 27° C. significantly interfered with subsequent Schiff reaction but did not affect the basophilia to any observable degree.

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Hypophyses of mice ranging from birth to 150 days of age of both sexes were examined in this experiment.
1. Acidophile cells which were found in only small numbers at immediately after birth showed a gradual increase in number and size with advancing age, and at about 30 days of age many acidophiles showed, though small in size, a similar appearance as seen in adults. At about 60 days of age the cells attained a fully developed state in cytological detail. The number of acidophiles indicated a slight de- crease at the age of150 days. On the other hand, as far as the aci- dophiles were concerned, there was found no significant difference be- tween both sexes throughout this experiment.
2. Two types of basophile cells could be distinguished. The delta-cell, which contains PAS positive, fine cytoplasmic granules, was situated mainly in the anterior portion of the anterior lobe and partly in the central area of the lobe, and the beta-cell which reacts positively with aldehyde-fuchsin as well as with PAS reagent was located in the central region of the lobe. The presence of the former was faintly recognized at a time immediately after birth, while the latter was almost not found till about 10 days of age. These two types became clearly distinguishable at about 20 to 30 days after birth.
3. At 40 to 50 days of age, marked sexual differences occurred in the delta-cell. In females at this stage delta-cells with hyaline vacuoles in the cytoplasm were encountered frequently, and their cytoplasmic granules decreased in number and size when compared with those at just before this stage. On the contrary, in males, such a phenomenon concerning delta-cells did not occur. Thus, in adults, delta-cells in males predominated those in females in number and size, although in males the cells with cytoplasmic hyaline vacuoles were far less in number than in females.
4. As far as the beta-cell was concerned, there was found no difference between both sexes throughout this experiment.
5. In females, a marked occurrence of the“ peculiar chromophobe cell” was observed from the 40th day on, although these cells were only rarely encountered in males throughout this experiment. The cells possessed an eccentrically situated, somewhat dark or vesicular nucleus and a cap-like structure consisting of many basophilic filaments at the cell periphery.
6. The increase in number of the acidophile cell depended mainly on mitotic division of the cell itself and partly on a transition from chromophobe cells. On the other hand, the number of basophile cells increased almost entirely by means of a transition from chromophobes.

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