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Effect of iodoacetic acid on ERG and retinal ultrastructure
~1 l:s~l’I:A(“~‘~
1 ‘ifi
Sympathetic ( ;. i,. RUSKELL.
Innervation
of the Ciliary
XIuscle in Monkeys
kI,fiiO,l.
The electron microscope was used to seek confirmat,ion of a sparse arlrener~g~c~ sytnlrathetio mnervation of the ciliary muscle observed recently with the formaldrhydr~ indured tl~~~rcscen~e technique. Small granular vesicles characteristic of sympathetic terminal varicositirs \\erc~ fLu1~1 ntijapent to muscle fibres in most but not all monkey ciliary muscles. Sympathetic t.erminals jvei’(‘ rc,att,erc!tl t.hroughout all regions of the muscle wit,11 no marked concentrations. Retwcen 1) and 2.,5”,, 01’ about 2000 terminals of each muscle were identified as sympathetic. In contrast half lhc tcrmiuals fount1 in arteriole walls within the muscle mere sympathetic. Terminals with small granular veai~~l~s w<‘nh not observed after superior cervical gsnglionectomy. The very small proportion and inconstancy of the sympathetic tctrmiaals suggcsti thic: t hc:g have a negligible role in ciliary mr~sclc control.
Determination
of Amino Acids in Vitreous Body of Single Human Eyes Xarburg
L. WELCE-LUSSES,
In the vitreous of two human healthy eyes, one cadaver eye and 13 pathological eyeballs t,hc concentration of free and bound amino acids was measured. Compared to two fresh vitreous bodies we found in the vitreous of a cadaver 15 hr post mortem a higher concentration of aspartic acid, glycine, alanine, phenylalanine and serine in free and bound form, also methionine in free form, aft,er hydrolysis also cysteic acid and ornithine. Jn four eyeballs affect’ed by melanoma, except one case with perforation through the sclera, the free amino acids differed less from healthy eyes than the amino acids in the bound form, especially aspartic and glutamic acids. In general absolute glaucomas show higher concentrations than acute glaucomas owing to an increase of the bound fraction. Causes of acute glaucoma showed hardly any ditrerences in amino-acid composition as compared with normal eyes. One eye, affected by phthisis, differed scarcely from the values of t,he normal eyes, but a child’s eyeball with phthisis showed very high values, especially for the amino acids in bound form.
The Fine Structure of the Vitreous Rhesus Monkey I?. J. RENTSCH,
and its Contact to the Inner Surface of the Eye in the
Tubinged
The inorganic dye ruthenium red which stains mucopolysaccharides was used to investigate the vitreous in the rhesus monkey by electron microscope. Special attention has been paid to the membrana hyaloidea, the vitreous base and the contact of the vitreous to the retinal surface. The anterior hyaloid membrane consists of cross-striated fibers (collagen) which are coated and mterconnected by ruthenium red positive material, possibly representing hyaluronic acid. Near the ora serrata the zonular fibers split completely into single fibrils which mix with the fibrils of the vitreous base. The fibrils are interconnected by a network of mucopolysaccharide chains, which near the inner surface of the eye come into close contact with the mucopolysaccharides of the basement membrane (membrana limitans interna). The basement membrane is rich in ruthenium red positive material closely related to the outer lamella of the cell membrane. Slso in the posterior parts of the eye the contact between the vitreous cortex and the retinal surface is maintained by mucopolysaccharide bridges connecting the basement membrane and the mucopolysaccharide coat of the vitreous fibrils. There is no direct connection between the vitreous or zonular fibrils and the basement membrane. This junction takes place rather by the ruthenium red positive material coating these fibrils and merging with the basement membrane material.
Effects of Iodoacetic J. BABEL
Acid on ERG and Retinal Ultrastructure
and K. STANGOS,
Ge?zeon
Iodoacetic acid was given by intravenous (20 and 30 “g/kg) and intravitreous (0.2 ml of a O.U15(!6 solution) injections to adult rabbits. Averaged ERG was recorded until complete or partial (flattening of b-wave) abolition occurred; the animals mere killed for EM-examination of retinas from 30 min up to 3 hr after injections. * This work J. 11”. Rohen.
ha,s been
done
in part
by the Anatomical
Institute
of Xarburg,
director
Prof.
Dr
ABSTRACTS
177
El% 4 11as abolished 2-7 min after intravenous injection whereas after int’ravitreous injection alterat,ions of ERG reached their maximum only 15-20 min after the injection, without. ever leading t,o a complete extinction of retinal activity. This diflerence of behaviour may be explained hv a faster raise of the iodoacetic acid concentration in the outer retinal layers after iutravenous injections. illterations of ERG showed up in a characteristic temporal order: a fast diminution of b-have amplitude preceding charges of a-wave, oscillary potentials and temporal parameters. The first detectable ultrastructural lesions were nresent in &Killer cells which showed distended and part,ly disrupted endoplasmic reticulum, interpreted as si,ss of altered metabolic activity of the glia. Oedema of optic nerve fibres occurred after intravitreous injection, but this is not implicated in the biogenesis of ERG. No other ultrastructural retinal lesions became evident within 3 h of the injection. The a hove observations lend support to the Jliiller cells being implicated in the biogenesis ot ERG. (Supported by SNSF, grant 3300.70.)
Retinal Glycogen in Relation to BIood Circulation D. KASAJ~L.
0. HO~KWIN,
U. METZLER
In the rabbit retinal glycogen, interruption of the blood supply
Contributions Retinas
and C. SCHEDLER,
which was to t’he eye.
traced
Ram
histochemically,
of Rods and Cones to the Electroretinographic
disappears
3045
min
Responses of Isolated
after
Human
Human retinas isolated in daylight respond to intense whim stimuli with photopic electroretinograms (Yickel et al., 1961). In the present study, evidence for a functioning scotopic system was obtained from human retinas dark adapted prior to, and during, the isolation procedure. The retinas were dissected from eyes enucleated because of malignant tumours. Retinal pieces of 0.3 cm* were incubated at 36°C in a glucose-containing ionic medium of pH 7.4. No plasma was added to the perfusate. Electroretinographic responses to monochromatic stimuli were recorded. Scotopic b-waves were obtained in 7 out of 8 retinas. The amount of energy required to elicit a just detectable response corresponded to approximately one quantum absorbed per rod when the stimulus lasted 30 msec. The threshold irradiance of longer stimuli (600 msec) amounted to 4 :; 1OF erg se+ cm-?, is no more than 3.6 log units above the corresponding psychophysical threshold. Similar weak stimuli were sufficient to elicit threshold P III responses in ageing preparations. Scotopic spectral sensitivities were found for both b-wave and P III. After exposure to intense radiation at 503 nm which bleached substantial amounts of rhodopsin, the eleetroretinogram consisted merely of components showing a photopic spectral sensitivity.
Siekel, \V., Lippmann, 63, 316-318.
Electron Microscopic Retina IV. LEIK;HE,
H. G., Haschke,
Observations
REFEREKCE \I-. and Baumann,
on Degeneration
Ch.
(1961).
Ber.
dtsch.
in the Pigment Epithelium
opldhd.
Ues.
of the Human
Hnnaburg
After a short explanation of the normal structure of t.he pigment epithelium of the human eye the author ctescribes the different changes within the cell which should be regarded as degenerative. The morphological changes consist of: (1) different sized vacuoles or cystic cavities which occur singly or in groups, spread out in the cvtoplasm or in the base of the cell. The endoplasmic reticulum shows small vacuoles instead o> tubules in cytoplasm. These vacuoles are found close to each other. (2) (iiant membrane structures which are lined up in terms of a circle or of an arch and at some places chains of vacuoles are to be detected; (3) conglomerations of long ergastoplasmic lamellae with terminal cystic enhancement,s; (4) lamellar systems looking like degenerate outer segments, and consisting of concentric layers of membranes ; (5) round homogenous and electron-dense bodies located in the cell, sometimes in direct contact to I hi: nucleus.
1 ‘ifi
Sympathetic ( ;. i,. RUSKELL.
Innervation
of the Ciliary
XIuscle in Monkeys
kI,fiiO,l.
The electron microscope was used to seek confirmat,ion of a sparse arlrener~g~c~ sytnlrathetio mnervation of the ciliary muscle observed recently with the formaldrhydr~ indured tl~~~rcscen~e technique. Small granular vesicles characteristic of sympathetic terminal varicositirs \\erc~ fLu1~1 ntijapent to muscle fibres in most but not all monkey ciliary muscles. Sympathetic t.erminals jvei’(‘ rc,att,erc!tl t.hroughout all regions of the muscle wit,11 no marked concentrations. Retwcen 1) and 2.,5”,, 01’ about 2000 terminals of each muscle were identified as sympathetic. In contrast half lhc tcrmiuals fount1 in arteriole walls within the muscle mere sympathetic. Terminals with small granular veai~~l~s w<‘nh not observed after superior cervical gsnglionectomy. The very small proportion and inconstancy of the sympathetic tctrmiaals suggcsti thic: t hc:g have a negligible role in ciliary mr~sclc control.
Determination
of Amino Acids in Vitreous Body of Single Human Eyes Xarburg
L. WELCE-LUSSES,
In the vitreous of two human healthy eyes, one cadaver eye and 13 pathological eyeballs t,hc concentration of free and bound amino acids was measured. Compared to two fresh vitreous bodies we found in the vitreous of a cadaver 15 hr post mortem a higher concentration of aspartic acid, glycine, alanine, phenylalanine and serine in free and bound form, also methionine in free form, aft,er hydrolysis also cysteic acid and ornithine. Jn four eyeballs affect’ed by melanoma, except one case with perforation through the sclera, the free amino acids differed less from healthy eyes than the amino acids in the bound form, especially aspartic and glutamic acids. In general absolute glaucomas show higher concentrations than acute glaucomas owing to an increase of the bound fraction. Causes of acute glaucoma showed hardly any ditrerences in amino-acid composition as compared with normal eyes. One eye, affected by phthisis, differed scarcely from the values of t,he normal eyes, but a child’s eyeball with phthisis showed very high values, especially for the amino acids in bound form.
The Fine Structure of the Vitreous Rhesus Monkey I?. J. RENTSCH,
and its Contact to the Inner Surface of the Eye in the
Tubinged
The inorganic dye ruthenium red which stains mucopolysaccharides was used to investigate the vitreous in the rhesus monkey by electron microscope. Special attention has been paid to the membrana hyaloidea, the vitreous base and the contact of the vitreous to the retinal surface. The anterior hyaloid membrane consists of cross-striated fibers (collagen) which are coated and mterconnected by ruthenium red positive material, possibly representing hyaluronic acid. Near the ora serrata the zonular fibers split completely into single fibrils which mix with the fibrils of the vitreous base. The fibrils are interconnected by a network of mucopolysaccharide chains, which near the inner surface of the eye come into close contact with the mucopolysaccharides of the basement membrane (membrana limitans interna). The basement membrane is rich in ruthenium red positive material closely related to the outer lamella of the cell membrane. Slso in the posterior parts of the eye the contact between the vitreous cortex and the retinal surface is maintained by mucopolysaccharide bridges connecting the basement membrane and the mucopolysaccharide coat of the vitreous fibrils. There is no direct connection between the vitreous or zonular fibrils and the basement membrane. This junction takes place rather by the ruthenium red positive material coating these fibrils and merging with the basement membrane material.
Effects of Iodoacetic J. BABEL
Acid on ERG and Retinal Ultrastructure
and K. STANGOS,
Ge?zeon
Iodoacetic acid was given by intravenous (20 and 30 “g/kg) and intravitreous (0.2 ml of a O.U15(!6 solution) injections to adult rabbits. Averaged ERG was recorded until complete or partial (flattening of b-wave) abolition occurred; the animals mere killed for EM-examination of retinas from 30 min up to 3 hr after injections. * This work J. 11”. Rohen.
ha,s been
done
in part
by the Anatomical
Institute
of Xarburg,
director
Prof.
Dr
ABSTRACTS
177
El% 4 11as abolished 2-7 min after intravenous injection whereas after int’ravitreous injection alterat,ions of ERG reached their maximum only 15-20 min after the injection, without. ever leading t,o a complete extinction of retinal activity. This diflerence of behaviour may be explained hv a faster raise of the iodoacetic acid concentration in the outer retinal layers after iutravenous injections. illterations of ERG showed up in a characteristic temporal order: a fast diminution of b-have amplitude preceding charges of a-wave, oscillary potentials and temporal parameters. The first detectable ultrastructural lesions were nresent in &Killer cells which showed distended and part,ly disrupted endoplasmic reticulum, interpreted as si,ss of altered metabolic activity of the glia. Oedema of optic nerve fibres occurred after intravitreous injection, but this is not implicated in the biogenesis of ERG. No other ultrastructural retinal lesions became evident within 3 h of the injection. The a hove observations lend support to the Jliiller cells being implicated in the biogenesis ot ERG. (Supported by SNSF, grant 3300.70.)
Retinal Glycogen in Relation to BIood Circulation D. KASAJ~L.
0. HO~KWIN,
U. METZLER
In the rabbit retinal glycogen, interruption of the blood supply
Contributions Retinas
and C. SCHEDLER,
which was to t’he eye.
traced
Ram
histochemically,
of Rods and Cones to the Electroretinographic
disappears
3045
min
Responses of Isolated
after
Human
Human retinas isolated in daylight respond to intense whim stimuli with photopic electroretinograms (Yickel et al., 1961). In the present study, evidence for a functioning scotopic system was obtained from human retinas dark adapted prior to, and during, the isolation procedure. The retinas were dissected from eyes enucleated because of malignant tumours. Retinal pieces of 0.3 cm* were incubated at 36°C in a glucose-containing ionic medium of pH 7.4. No plasma was added to the perfusate. Electroretinographic responses to monochromatic stimuli were recorded. Scotopic b-waves were obtained in 7 out of 8 retinas. The amount of energy required to elicit a just detectable response corresponded to approximately one quantum absorbed per rod when the stimulus lasted 30 msec. The threshold irradiance of longer stimuli (600 msec) amounted to 4 :; 1OF erg se+ cm-?, is no more than 3.6 log units above the corresponding psychophysical threshold. Similar weak stimuli were sufficient to elicit threshold P III responses in ageing preparations. Scotopic spectral sensitivities were found for both b-wave and P III. After exposure to intense radiation at 503 nm which bleached substantial amounts of rhodopsin, the eleetroretinogram consisted merely of components showing a photopic spectral sensitivity.
Siekel, \V., Lippmann, 63, 316-318.
Electron Microscopic Retina IV. LEIK;HE,
H. G., Haschke,
Observations
REFEREKCE \I-. and Baumann,
on Degeneration
Ch.
(1961).
Ber.
dtsch.
in the Pigment Epithelium
opldhd.
Ues.
of the Human
Hnnaburg
After a short explanation of the normal structure of t.he pigment epithelium of the human eye the author ctescribes the different changes within the cell which should be regarded as degenerative. The morphological changes consist of: (1) different sized vacuoles or cystic cavities which occur singly or in groups, spread out in the cvtoplasm or in the base of the cell. The endoplasmic reticulum shows small vacuoles instead o> tubules in cytoplasm. These vacuoles are found close to each other. (2) (iiant membrane structures which are lined up in terms of a circle or of an arch and at some places chains of vacuoles are to be detected; (3) conglomerations of long ergastoplasmic lamellae with terminal cystic enhancement,s; (4) lamellar systems looking like degenerate outer segments, and consisting of concentric layers of membranes ; (5) round homogenous and electron-dense bodies located in the cell, sometimes in direct contact to I hi: nucleus.