Cites M. Nussbaum, 'Ueber den Bau und die Thätigkeit der Drüsen: Die Fermentbildung in den Drüsen', Archiv für mikroskopische Anatomie 13 (1) (1877), pp. 721-755.
Description:

'Nussbaum [note: 'Nussbaum. Arch. f. Mik. Anat. Bd. XIII. p. 746, 1877.'] has described the appearance of the oesophageal glands after treatment with osmic acid. It will have been seen that we differ from him in many points.

1. He finds that after "several days' hunger" the alveoli are like the alveoli of the pancreas and present two zones nearly equal. We find that frogs killed four days after feeding present alveoli granular throughout. In two or three days after feeding some frogs do not completely recover their granules, possibly Nussbaum may have taken some such slowly reacting frogs, as the typically hungry. Possibly in some frogs the granular zone does not in the ordinary hungry condition stretch quite to the periphery; in the healthy frogs which we have observed it did, nevertheless the matter does not seeim to us very important.

2. On feeding, he finds the granules increase so that in three hours the alveoli are granular throughout, an appearance which they preserve until about five hours after feeding, and only at about the fifteenth hour has the outer clear zone again become obvious. We find that from the beginning of digestion onwards there is a using up of granules.

3. According to him, if small pieces of cork be given to a frog and the cardia ligatured so that the cork continues to stimulate the oesophageal mucous membrane, the granules disappear entirely in three to four hours from the alveolar cells, whilst the nuclei become shrunken and angular.

This we have been quite unable to observe.

In the granular throughout stage of Nussbaumn after five hours feeding, he describes coarser particles staining deeply black with osmic acid, immediately underneath the membrana propria. We have described in a similar position collections of small highly refracting granuiles which stain very deeply with osmic acid; these "border" granules perhaps represent the coarser particles of Nussbaum.

That which we regard as the most important point of difference is the effect of feeding: Nussbaum finds that the granules increase at first, we find only a decrease.' (285)

 

'Nussbaum connected the presence of granules in the oesophageal glands with the presence of ferment. His starting point was, however, not so much the actual presence of granules as their staining with osmic acid; according, to him, when the granules disappeared, the blackening from osmic acid ceased. This seems to us untenable, for if the gland be treated with osmic acid alone for two hours, the granules are not more deeply stained than the rest of the protoplasmic cells; whilst the border-granules are deep black, so that on Nussbaum's view, it should be these and not the central granules which would represent the ferment. We may remark too that Nussbaum's observations on the granularity of the cells do not with any accuracy correspond with the results of Swieçicki on the amount of ferment. The former finds the granules in the cells increase for three hours after feeding, remain stationary for two hours and then diminish. The latter found the ferment increase up to 6-10 hours and only then diminish.

From general reasoning we are inclined to connect the number of central granules with the amount of ferment, and we rnay point out that Grützner's [note: 'Grützner. Pflüger's Arch. Bd. XVI. p. 122, 1877.']  results agree thoroughly with this view. He found that the ferment was greatest during hunger, and diminished during digestion, at first rapidly, then more slowly.' (286)

 

'The greater curvature contains more border-cells than any other portion of the stomach, the pyloric glands of the smaller curvature contain at most an occasional border-cell here and there, yet the amount of pepsin produced by the two gland-forms is scarcely different. The conclusion is irresistible that the border-cells do not form the ferment.' [re: Nussbaum's assertion to the contrary. No direct reference to Nussbaum here.] (295)

 

'The granules we consider as stored up cell-products, which, on suffering molecular re-arrangement during the secretion, give rise amongst other substances to the proteolytic ferment.

We cannot agree with Nussbaum's view that the depth of staining with osmic acid is a trustworthy index of the amount of ferment present in the cells. On his view, it appears to us, the border, rather than the central, granules should be connected with the ferment.' (297-298)

 

Cites Plate IX, Journal of Physiology 2 (4) (1879). Figs. 1-15 and A from J.N. Langley and H. Sewall, 'On the Changes in Pepsin-forming Glands During Secretion'.
Description:

Explanation of Plate IX:

'Fig. 1-4. Oesophageal gland of Frog in the living state. Outlines drawn with Camera oc..3, obj. A, Zeiss.

Fig. 1. Edge of a lobule from a frog four days after feeding.

Fig. 2. 11 hours after feeding.

Fig. 3. 6 hours after feeding.

Fig. 4. From an unhealthy frog. Shows the clumps of border granules. This frog had been fed the day previous to examination; much the same appearance is, however, often shown by unhealthy frogs two, three or more days after feeding.

Figs. 5, 6. Oesophageal Gland of Frog treated with osmic acid and subsequently with alcohol. Outlines drawn with Camera oc. 3, obj. 7, Hartnack.

Fig. 5. 4 days after feeding.

Fig. 6. 5 hours after feeding a four-days hungry frog.

Figs. 7-10. Gastric Glands of Triton toeniatus in fresh state.

Fig. 7. 24 hours after feeding with worm.

Fig. 8. 3 hours-after feeding with worm. The three glands drawn show the different extent to which glands of the same stomach may be affected.

Fig. 9. 6 days after feedilng.

Fig. 10. 40 minutes after introduction of albumin into exposed stomach. For method cp. p. 288. Viewed longitudinally. The change was in this case more rapid than usual. The nuclei to the right of the figure have too distinct an outline.

Figs. 11, 12. Gastric Glands of the Stickleback in the fresh state.

Fig. 11. Hunger.

Fig. 12. 3 1/2 hours after feeding with worm.

Fig. 13. Body of gastric gland of Cat. Taken from the larger curvature near the fundus. In fresh state. Camera oc. 3, obj. 7, Hartnack.

Fig. 14. Neck of similar gland.

In both the granules mark the chief-cells, the lightly shaded parts the border-cells. In Fig. 14 the isolated patches of granules indicate the chief-cells of the neck of the glands occurring amidst a mass of border-cells. A very slight trace of acid makes the border-cells distinctly granular. Camera oc. 3, obj. 7, Hartnack.

Fig. 15. Transverse section of glands of smaller curvature of rabbit in the fresh state. Camera oc. 3, obj. 7, Hartnack.

Fig. A. Rabbit's stomach; to show regions in which different gland-forms occur. Life-size.

1. Fundus. Border-cells and coarsely granular chief-cells.

2. Larger curvature. Border-cells and chief-cells not distinctly granular.

3. Smaller curvature and region around pylorus. Ordinary pyloric glands, no cell distinctly granular.

4. Pylorus. The region between 1 and 2 shifts markedly in character at different times: in hunger becoming more like 1, in digestion more like 2.' (300-301)

 

Figs. 1-3 in text:

'The oesophageal glands of a frog three to four days after such a meal [of worms] show alveoli densely and darkly granular, no cell-outline is visible, and the outlines of the separate alveoli are but faintly indicated (PI. IX. Fig. 1).

There is no marked distinction between the mucous-cells and those of the rest of the alveoli.

If to such a hungry frog a worm be given, the glands begin almost immediately to change in appearance. The rapidity of onset of this change varies considerably in different frogs, in an hour it is usually marked. The alveoli are no longer granular throughout, but have an outer clear non-granular zone and an inner zone adjacent to the lumen thickly granular as before. The alveolar outlines are more distinctly though still faintly marked, nuclei are not visible any more than in the hungry specimen. The time of most rapid change is the first few hours (Fig. 2), later it proceeds slowly. After about five hours the granules continue to disappear, but they disappear not so much at the junction of the clear and granular zones as throughout the latter. Thus the inner zone becomes less granular and less dark, but the outer clear zone does not increase to any marked degree. At the same time the granules take an irregular star-shape, the clear zone no longer being an even rim, but here and there dipping more deeply down towards the lumen (Fig. 3).' (282-283)

 

Fig. 4 in text:

'Besides the above-mentioned granules, the "central" granules, there are others smaller and somewhat more refractive, which from their position we may call "border" granules; they are usually arranged in clumps immediately uinderneath the basement membrane (Fig. 4). These were not noticed in the earlier used and most healthy frogs, they may however have been present.' (283)

 

Figs. 5 and 6 in text:

'The results spoken of above as obtainable in the fresh state, are also obtainable by treating the gland with osmic acid; the living appearances are in the main preserved. The clear zone is however more conspicuous and the star-arrangement of granules more marked in the hardened specimens; this is in part due to the action of the acid on the granules in the mucous-cells, and in part to the smaller thickness of cell-substance looked at. The outlines of the cells are but faint, the nuclei only to be made out with difficulty, whilst the nucleoli come out sharply. The granules are somewhat larger. The border-granules, when present, are the most deeply-stained constituent of the alveolar cells. The mucous-cells are conspicuous, being no longer granular but clear and transparent, the acid either dissolves out the granules or causes them to run together, so as to be indistinguishable. The mucous-cells, then, show a small mass of stained protoplasm containing an irregular nucleus, from which a network, such as is usually seen in mucous-cells, stretches out (compare Figs. 5 and 6).' (284)

 

Figs. 7 and 8 in text:

'Only those observations were depended on in which there was good reason to suppose that the vital conditions were fairly normal. To induce digestive changes either a worm was given to the newt, or some dilute egg-albumiin introduced into the stomach. The gastric glands of a newt thus observed twenty-four hours after feeding the animal, are densely and somewhat finely granular throughout, the separate cells are not to be distinguished and no nuclei are visible (Fig. 7).

If taken three hours after feeding there are markedly fewer granules; the different glands however vary widely. In some few, scarcely any granules are to be seen, in others they are collected in a band round the lumen, leaving a more or less ill-defined outer clear zone; some glands still have granules throughout. Usually there are many glands in which the cells are to some extent defined; in these the granules are commonly collected along the lateral margin of the cells and near the lumina, forming thus a star-like figure (Fig. 8).' (287)

 

Fig. 9 in text:

'If the newt be kept without food longer than one day, the granules appear slowly to diminish, so that after six or more days the description given above of the digesting stomach begins to some extent to apply, i.e. the granules thin away at the periphery (Fig. 9). To what further extent this proceeds we have made no experiments to determine.;' (287-288)

 

Fig. 10 in text:

'The course of events is usually as follows. In each gland within about half-an-hour after mounting, near the middle of the gland cells corresponding to the position of the nuclei, ill-defined round clear spots make their appearance. These nuclear spots increase in size so that the granules become arranged as a mesh in the gland; then in some the granules disappear at the outer part of the cells giving the star figure (Fig. 10). In many of these latter, the granules disappear from the branches of the star, ie. around the nuclei, and give ultimately the inner granular and outer clear zone. In this last stage, occurring usually in three to five hours, the distinction between the two zones is commonly more striking than when viewed by the first method.' (289)

 

Figs. 11 and 12 in text:

'The gastric glands of a stickleback, which has hungered three to six days, are granular throughout, somewhat finely, the granules thinning away more or less from the centre to the periphery (Fig. 11). The lumina are inconspicuous, and the glands of fairly equal size.

In about three to five hours after feeding the animal with a worm, a very marked change is seen in the glands. The granules, become perhaps rather larger, are aggregated round the lumen, leaving usually a distinct peripheral rim. The cells are smaller, and the lumen often increased to such an extent as to suggest that the inner margins of the cells have been broken down in the process of secretion; fig. 12 is taken from a stomach three hours after feeding. Although this is the prevailing appearance of the digesting stomach, yet there are usually some glands approximating more or less to the hungry state. ' (290-291)

 

Figs. 13-15 in text:

'In all mammals which we have observed the chief-cells are thickly and coarsely granular (Fig. 13), and the pyloric cells are by comparison transparent, and very finely granular, often appearing homogeneous (Fig. 13). It is known that in the necks of the fundus-glands, here and there, chief-cells occur amongst the border-cells; in the fresh state small patches of dark granules are often seen, which on treatment with reagents turn out to be chief-cells (Fig. 14).

The above described living condition of the chief-cells is characteristic of the hungry animal; but just as, in activity, the granules diminish in many serous glands of mammals and pepsin-forming glands of other animals; so, in digestion, do the granules diminish in the chief-cells of the mammals' stomach.' (292)

 

Fig. A in text:

'The rabbit presents greater difficulties for examination than the cat or rat, since the mucous membrane varies in structure, almost from point to point. It may be conveniently divided into four parts, the typical glands of each part however graduating into the neighbouring ones (Fig. A).

1. The Fundus is to the unaided eye of an opaque white on the surface turned to the muscle-coat. The glands contain comparatively few border-cells, the greater part of the gland consists of darkly granular chief-cells. In approaching the greater curvature the border-cells become more numerous, and the chief-cells more confined to the deeper portions of the glands; at the same time the granules become fewer and appear in transverse section of the glands as an irregular star.

2. The median part of the Greater Curvature. This is pink and contrasts strongly with the fundus. In fresh specimens scarcely any dark chief-cell granules are to be seen, there may be a few scattered irregularly in the central portion of a gland, the rest of the gland being finely granular, without cell outlines, except where the cells bulge at the outer border.

3. The Smaller Curvature and mucous membrane immediately around the pylorus. It is greyish-white, the glands here are what are usually described as pyloric glands.

4. The Pylorus. The glands are rather shallow depressions of the mucous membrane, the cells are clear, more transparent than the pyloric gland-cells.' (293)

Cited by J.N. Langley, 'On the Histology and Physiology of Pepsin-forming Glands', Philosophical Transactions of the Royal Society of London 171 (1881), pp. 663-711.
Description:

'Sewall and myself [note: 'Langley and Sewall, Proc. Roy. Soc., Oct., 1879, p. 383; Jour. of Physiol., vol. ii., p. 283, 879.'] found that the oesophageal gland-cells occasionally showed clumps of highly refractive granules in their peripheral portions. To distinguish these from the proper granules of the cells, we called them "border" granules. Further investigation has shown me that these are really fat globules. Their position is very constant and they give a striking appearance to osmic acid specimens. Quite similar fat globules are occasionally to be seen in the pancreatic gland-cells. They occur also in the gastric gland cells of the Frog, Toad, and Newt (see Plate 78, fig. 7), although their arrangement is not quite so regular. In a subsequent paper I hope to discuss the causes which influence the appearance of fat globules in the above and in other secretory gland-cells.' (665)

 

'It was stated by Sewall and myself [note: 'Op. cit., p. 283'] that absolute alcohol added to the fresh teased-out gland altered the normal appearances; and I have said above that the granules are in part dissolved. Nevertheless, alcohol specimens of oesophageal glands taken during digestion show the two zones in the gland-cells; the non-granular zone stains with carmine, and thus specimens can be obtained (Plate 77, fig. 8), which, except for the smaller size of the cells, closely resemble similarly prepared specimens of the pancreas. It is almost unnecessary to remark that the "granules" of the inner zone in alcohol specimens are not the granules present in the fresh gland. With Zeiss', oc. 2, obj. E or F, the granular zone appears as a fine network.' (670)

 

'The extent to which the disappearance of granules proceeds varies in different cases: in many cases two days after feeding [note: 'Nussbaum (op. cit., s. 749) made some observations upon the direct stimulation of the oesophageal mucous membrane, the cardia being ligatured before the animal was fed with sponge. He found under such circumstances that the granules entirely disappeared from the cells in three to five hours. Sewall and myself (op. cit., p. 285) were unable to observe any such rapid action.'] with a rather large piece of sponge, occasionally in a less time, scarcely any granules are left; and in some glands not a granule is to be seen (Plate 77, fig. 6 (a), 6 (b)).' (671)

 

'In the account of the changes in the oesophageal glands given by Sewall and myself there were two points which clashed with the earlier observations which Nussbaum made on osmic acid specimens. He described the gland-cells of the normal hungry Frog as having a large clear zone, and found that on feeding the animal the granules increased so that in three to five hours a clear zone was no longer to be seen.

Grutzner's [note: 'Grützner, Pflüger's Arch., Bd. xx. s. 395, 1879.'] results suggest an explanation of the divergence between the account of Nussbaum and that of Sewall and myself. He finds that in the normally hungry Frog the oesophageal glands are granular throughout and diminish in granularity during digestion; but finds also that if a Frog is kept longer than usual without food a clear zone is then formed in the oesophageal glands and that on feeding there is at first an increase of granules. Thus according to Grützner, Nussbaum's results would represent what occurs in a pathological and not what occurs in a normal condition,

During the last year and a half I have made a considerable number of observations with the view of determining the points at issue. As regards the state of the glands in the normal hungry Frog I have seen no reason to alter my first-formed opinion.

In some Frogs a large clear zone in the oesophageal glands does occur, namely, in those in which there are signs of general inflammation. In nearly all cases in which I have found a marked clear zone in the gland-cells of a hungry Frog, the animal had some mark or other of an ill state of health. I have frequently selected lively, active Frogs, and sluggish, unhealthy ones from a batch brought to the laboratory, and in a few days examined the oesophageal glands. The gland-cells in the former were granular throughout; those of the latter had almost always a clear zone.' (672)