The distribution of neuropeptides in the ocular tissues of several mammals: A comparative study

1. The distribution of several neuropeptides (vasoactive intestinal peptide, substance P, somatostatin and neurotensin) was assessed in ocular tissues from the cow, sheep, rabbit and rat.2. Vasoactive intestinal peptide was most abundant in the choroid and sciera in all species except the rat. Substance P was most abundant in the retina of cow and rat and in the iris/ciliary body of sheep and rabbit. Somatostatin and neurotensin were most abundant in the retina of all species examined.3. Regulatory peptides thus display distinct regional distributions within the ocular tissues of a single species of mammal and, in addition, exhibit interspecific variation.     

Effect of anti-pig zona pellucida serum on fertilization in several mammals

The effect of goat antiserum against isolated pig zonae pellucidae on fertilization in vivo was examined in the pig, cow, sheep, rabbit, rat, and mouse. As shown by indirect immunofluorescence, anti-pig zona serum reacted strongly with the zonae of pig, cow, sheep, and rabbit, but the reaction with the zonae of mouse and rat was weak. Passive immunization with anti-pig zona serum significantly, or completely, inhibited fertilization in all species. However, inhibition of fertilization was more pronounced in the pig, cow, sheep, rabbit, and mouse than in the rat. Inhibition of fertilization in the rabbit was also observed after passive immunization with antiserum absorbed with rabbit liver and kidney. All of the zonae recovered from the pig, cow, sheep, rat, and mouse after passive immunization with anti-pig zona serum exhibited strong fluorescence, regardless of the incidence of fertilization. It was concluded that the pig and other mammalian zonae pellucidae tested have tissue-specific antigens.     

Studies on hormonal regulation of lipolysis and lipogenesis in fat cells of various mammalian species

1. Corticotropin-stimulated lipolysis in adipocytes of rats, mice, hamsters, guinea pigs and rabbits. Melanotropins elicited high lipolytic activity only in guinea pig and rabbit adipocytes. Opiate peptides were active only in rabbit adipocytes. Pituitary and chorionic gonadotropins and somatotropin were lipolytic in guinea pig adipocytes. Other hormones tested including prolactin, somatostatin, substance P, neurotensin, angiotensin II, thyrotropin releasing hormone and pancreatic polypeptide were devoid of lipolytic activity in all of the adipocytes studied. 2. In the rabbit adipocytes gamma-melanotropin was lipolytic only at high doses. At these doses the peptide inhibited the lipolytic response to a high dose of corticotropin. 3. Lipolysis stimulated by vasoactive intestinal peptide and epinephrine in rat adipocytes was antagonized by insulin. The lipolytic hormones corticotropin, epinephrine, vasoactive intestinal peptide and secretin suppressed basal and insulin-stimulated lipogenesis.     

Taurine and other free amino acids in the retina,vitreous, lens,irisciliary body,and cornea of the rat eye

Levels of free amino acids were determined quantitatively in whole ocular tissues of the rat eye with aid of a sensitive amino acid analyzer. The tissues studied were the retina, vitreous, lens, iris-ciliary body, and cornea. The retina and lens contained a more concentrated free amino acid pool than other tissues. The neuroactive amino acids taurine, GABA, glutamic acid, aspartic acid, and glycine were clearly enriched in the retina. Taurine was the most abundant amino acid in all five tissue studied, and its high concentration in non-neural tissues, especially the lens, suggests that it must have other functions as well as neurotransmitter ones in the rat eye.     

Species and organ dependence of alkaline phosphatase activity in lymphatic tissues. A histochemical, biochemical and electrophoretic study

Synopsis Alkaline phosphatase activity in lymphatic tissues of guineapig, cat, cow, dog, rabbit, sheep, rat, mouse, hamster, chicken and man was studied with histochemical, biochemical and electrophoretic techniques. The thymus showed decreasing alkaline phosphatase activity from species to species in the order just given. Activity of alkaline phosphatase in other lymphatic tissues did not show such clear species and organ dependence. Spleens of the cat, cow and rabbit and lymph nodes of the cow and sheep gave, however, very characteristic patterns of alkaline phosphatase activity. In the chicken there was no difference between the alkaline phosphatase content of the thymus and that of the bursa of Fabricius. The lymphatic follicles of human tonsils and appendix and in the appendix of the rabbit exhibited alkaline phosphatase activity in the circular cell layer. This was also seen in some follicles in the lymph nodes of certain species. Electrophoretically, the main alkaline phosphatase fraction of the lymphatic tissues closely resembled the main fraction of blood, though it is probably not identical with it. Although the biological function of alkaline phosphatase is unknown, the greatly varying alkaline phosphatase content in different lymphatic organs of different species indicates that immunological studies with one species or with cells derived from a certain lymphatic tissue or with both are probably not directly comparable with studies using other species or cells from other lymphatic tissues.     

Somatostatin and VIP neurons in the retina of different species

Neurons displaying somatostatin or vasoactive intestinal polypeptide (VIP) immunoreactivity were detected among the amacrine cells in the retina of baboon, cynomolgus monkey, squirrel monkey, cow, pig, cat, rabbit, guinea-pig, rat, mouse, frog and goldfish. Generally, immunoreactive cell bodies were located in the inner nuclear layer with processes ramifying in three more or less well-defined sublayers in the inner plexiform layer. The density of the sublayers and their location varied with the peptide and species investigated. In most cases there was a sublayer in the outermost part (Ramon y Cajal's sublamina 1) of the inner plexiform layer and this sublayer was usually the best developed. In some species a few somatostatin fibres were also detected in the outer plexiform layer, suggesting that some interplexiform cells contain somatostatin. In the baboon VIP was found exclusively in interstitial amacrine cells which have their cell bodies and processes entirely within the inner plexiform layer.     

Quantitation and immunohistochemistry of catecholamines in the posterior segment of the eye

Summary Dopamine, noradrenaline, and adrenaline were assayed with HPLC in the light adapted retinae of carp, frog, chicken, pigeon, rat, guinea-pig, rabbit, cat, pig and cow. Dopamine varied from 0.6 to 2.6 nmol/g wet weight and was not influenced by sympathectomy. The dopamine figures agree with previously published results. Noradrenaline concentrations varied from not detectable to 0.06 nmol/g wet weight in different species. Homolateral sympathectomy significantly decreased the noradrenaline figure in rabbits. There are no previous figures for noradrenaline for most of the species. Adrenaline was not detected in any species. Immunohistochemical analysis showed noradrenaline to be present in choroidal nerves, but noradrenaline immuno-reactivity was not seen in the retina (chicken, rat, guinea-pig, rabbit, cat, cow). It is concluded that dopamine is the major catecholamine in the retina. Noradrenaline was found present only in minute amounts in the assays, and much of its was likely to stem from sympathetic nerve fibres. The study did not demonstrate any noradrenergic neurons in the retina.     

Quantitation and immunohistochemistry of catecholamines in the posterior segment of the eye

Dopamine, noradrenaline, and adrenaline were assayed with HPLC in the light adapted retinae of carp, frog, chicken, pigeon, rat, guinea-pig, rabbit, cat, pig and cow. Dopamine varied from 0.6 to 2.6 nmol/g wet weight and was not influenced by sympathectomy. The dopamine figures agree with previously published results. Noradrenaline concentrations varied from not detectable to 0.06 nmol/g wet weight in different species. Homolateral sympathectomy significantly decreased the noradrenaline figure in rabbits. There are no previous figures for noradrenaline for most of the species. Adrenaline was not detected in any species. Immunohistochemical analysis showed noradrenaline to be present in choroidal nerves, but noradrenaline immuno-reactivity was not seen in the retina (chicken, rat, guinea-pig, rabbit, cat, cow). It is concluded that dopamine is the major catecholamine in the retina. Noradrenaline was found present only in minute amounts in the assays, and much of its was likely to stem from sympathetic nerve fibres. The study did not demonstrate any noradrenergic neurons in the retina.     

The intermediate filament complement of the retina: a comparison between different mammalian species

We compared the intermediate filament expression of the various cell types in the fully differentiated neural retina from rat, mouse, rabbit, guinea pig, cow, pig, and cat. Many cell types had an intermediate filament complement conserved across species boundaries, such as Müller cells and retinal ganglion cells. In some species (rabbit, guinea pig, and cow), however, we were unable to visualize GFA (glial fibrillary acidic)-positive retinal astrocytes, although such profiles were clearly visible in the remainder. Horizontal cell staining proved to be extremely species-variable. In rat and mouse the processes of these cells were identically displayed with antibodies to vimentin and all three neurofilament triplet proteins. In cow they decorated with antibodies to vimentin and antibodies to the two lower molecular weight neurofilament proteins alone, whereas in pig, rabbit and guinea pig all three neurofilament proteins but not vimentin were present. Finally cat horizontal cells stained for all three neurofilament proteins, some finer processes being additionally stainable with vimentin. A further surprise was the visualization of profiles positive only for the two lower molecular weight neurofilament proteins in the inner nuclear layer of both rabbit and guinea pig retina but not the other species. The implications of these results will be discussed.     

Localization of neurotensin-immunoreactive cells in the small intestine of man and various mammals.

Antibodies against synthetic bovine neurotensin were raised in rabbits and used to demonstrate neurotensin-immunreactive cells by immunohistochemical methods. In the jejunum and ileum of all species investigated (man, dog, monkey, cat, rabbit, sheep, rat, mouse, hamster, chinese hamster, gerbil, pig and guinea pig) cells were present in the mucosa, which reacted specifically with antineurotensin serum using the indirect immunofluorescence and peroxidase-antiperoxidase methods. In the monkey Tupaia the distribution of neurotensin-immunoreactive cells was examined by investigating serial sections through the entire gastro-entero-pancreatic (GEP) endocrine system, again showing most neurotensin-immunoreactive cells in the jejunum and ileum. The functional role of the presence of neurotensin immunoreactivity in the gut is discussed.     

Glucose 1,6-diphosphate in the erythrocytes of various species of mammal

Red blood cells from human, pig, cow, rabbit, rat and sheep were investigated for the occurrence of phosphoglucomutase multiple forms, G1,6P2 level, PGM, PRM and G1,6P2 synthetic activities. In all cases a species specific pattern of PGM isoenzymes was detected by starch gel electrophoresis. G1,6P2 mean values range from 33 to 122 nmol/ml RBC for pig and rabbit erythrocytes, respectively. The study of the correlation between the biphosphate content and the occurrence of the three measured enzymatic activities indicates a possible role of PRM activity (property of PGM2 isoenzymes) in the erythrocytic G1,6P2 catabolism.     

Localization of neurotensin-immunoreactive cells in the small intestine of man and various mammals

Summary Antibodies against synthetic bovine neurotensin were raised in rabbits and used to demonstrate neurotensin-immunreactive cells by immunohistochemical methods. In the jejunum and ileum of all species investigated (man, dog, monkey, cat, rabbit, sheep, rat, mouse, hamster, chinese hamster, gerbil, pig and guinea pig) cells were present in the mucosa, which reacted specifically with antineurotensin serum using the indirect immunofluorescence and peroxidase-antiperoxidase methods. In the monkey Tupaia the distribution of neurotensin-immunoreactive cells was examined by investigating serial sections through the entire gastro-entero-pancreatic (GEP) endocrine system, again showing most neurotensin-immunoreactive cells in the jejunum and ileum. The functional role of the presence of neurotensin immunoreactivity in the gut is discussed.Supported by the German Research Foundation     

Ontogeny of the distribution and colocalization of calbindin D28K within neural and endocrine cells of the gastrointestinal tract of fetal and neonatal sheep.

Using immunocytochemical techniques we have demonstrated that Calbindin D28K (CaBP) is present in the gastrointestinal tract of ovine fetuses early in development (by day 45). At day 45, CaBP was limited to neuronal elements in the developing intestine. By day 100, CaBP immunoreactivity was abundant in both epithelial endocrine cells and nerves of the submucous and myenteric ganglia. The location of CaBP containing cells and fibers was similar in duodenal sections taken from day 100 and term (145 days), as well as those taken from 24-48 h postnatal lambs. CaBP is colocalized in endocrine cells containing gastrin, glucagon, somatostatin and neurotensin, but not glucose dependent insulinotrophic peptide (GIP). Furthermore, it is extensively colocalized in nerve fibers and cells containing neurotensin but not somatostatin or vasoactive intestinal peptide. The colocalization of CaBP within various endocrine and nerve cells does not change in fetal sheep over the last one-third of gestation and there is no difference between fetal and neonatal sheep.     

Peptide-containing nerve fibers in the circumvallate papillae.

The occurrence and distribution of an array of neuropeptides and dopamine-beta-hydroxylase in the circumvallate papillae of monkey, pig, cow, ferret, cat, rat and mouse was studied by immunocytochemistry. The animals were chosen to represent species with different diets. Substance P/neurokinin A- and calcitonin gene-related peptide-containing fibers were numerous in the circumvallate papillae of all animals examined, with the highest frequency in monkey, pig, cow, rat and mouse; in ferret and cat moderate numbers were detected. Vasoactive intestinal peptide/peptide histidine isoleucine amide-containing fibers were numerous in the circumvallate papillae of pig, while they were moderate in number in monkey, ferret and mouse. Neuropeptide Y-containing fibers were few to moderate in number in the circumvallate papillae of all species. Galanin-containing fibers were numerous in the pig circumvallate papillae, while only a few fibers could be detected in monkey, cow, cat, rat and mouse. Somatostatin-containing fibers were seen only in the cat circumvallate papillae, gastrin-releasing peptide-containing fibers in the cow and cat, cholecystokinin/gastrin-containing fibers in the pig and cow. Dopamine-beta-hydroxylase-containing fibers were detected in all animals studied. They were few to moderate in number in the circumvallate papillae. There was no obvious link between the peptidergic innervation pattern and the food habits.     

Immunohistochemical localization of neurotensin in endocrine cells of the gut

Summary Endocrine cells displaying neurotensin immunoreactivity are found scattered in the jejuno-ileum of all mammals studied, including man. They are rather scarce in rat, guinea pig, rabbit and pig and fairly numerous in cat, dog and man. In most mammals the neurotensin cells predominate on the villi. Only in the dog are they more numerous in the crypts. In the chicken, neurotensin cells occur all along the intestinal tract. They are particularly numerous in the zone that joins the gizzard with the duodenum. The ontogeny of the neurotensin cells in the gut was studied in rats and chickens. In the rat, the cells are first observed in the jejuno-ileum immediately before birth. The adult frequency is reached 4–5 days later. In the chicken, neurotensin cells first appear in the colon in the 18 day old embryo and in the small intestine two days later (i.e. one or two days before hatching). A few days after hatching, the gut has achieved the adult number of neurotensin cells per unit area.     

Comparative studies on the gastric glycopeptide in eleven animal species.

1. Glycopeptides in the stomachs of eleven mammalian species, including human, rabbit, horse, cow, pig, goat, sheep, dog, cat, guinea pig and rat were assayed by determining the carbohydrate content of materials which remained after proteolysis. 2. The glycopeptide content was higher in the mucosa than in the muscular layer including serosa, especially in the porcine stomach and the fourth stomachs of the ruminants than in the stomachs of any other animals. 3. The glycopeptide, which was stained with both alcian blue and PAS, was absent or sparingly present in the mucosae of the human, rabbit, horse stomachs and in the mucosae of the first to third stomachs of the cow, goat and sheep, whereas in the mucosae of the pig, dog, cat, guinea pig and rat stomachs and in the mucosae of the fourth stomachs of the cow, goat and sheep, it was found in noticeable extents.     

Glucose-1,6-P2 synthesis, phosphoglucomutase and phosphoribomutase correlate with glucose-1,6-P2 concentration in mammals red blood cells

Glucose 1,6-biphosphate (G1,6P2) was measured in human, pig, cow, rabbit, rat and sheep red blood cells. Mean values are variable among the species and range from 33 to 122 nmol/ml RBC for pig and rabbit erythrocytes, respectively. The activities of G1,6P2 synthase, phosphoglucomutase (PGM) and phosphoribomutase (PRM) have also been assayed in red cell haemolysates of the same species. The correlations between the biphosphate content and the occurrence of the three enzymatic activities have been studied in order to gain an insight into the regulation of the G1,6P2 turnover in mammalian erythrocytes.     

Comparative studies of the protein composition of red blood cell membranes from eight mammalian species

The polypeptide pattern of red blood cell (RBC) membranes from cow, sheep, horse, rabbit, guinea pig, rat, mouse, analyzed by polyacrylamide gel electrophoresis, was compared to human RBC counterpart. Some qualitative and quantitative differences were noted. Among the high molecular weight components the bands 2.1- 2.3 appeared slightly decreased in rabbit and rat and increased in sheep RBC membranes. Band 3 appeared to have a higher molecular weight in the cow, guinea pig and mouse RBCs, and a lower molecular weight in the sheep RBCs. Band 4.1 from the RBC membranes of cow, sheep, rabbit and guinea pig was splitted into two sub-bands, while band 4.2 overlapped with band 4.1 in horse and guinea pig RBC membranes. There are marked differences in the number and position of bands in the 4.5 region, while band 4.9 is present in higher amounts in horse, rabbit and guinea pig RBC membranes. Band 6 (glyceraldehyde 3-phosphate dehydrogenase) was undetectable in horse, rat and mouse RBC membranes and was decreased in sheep, rabbit and guinea pig. There are also major differences in the region of band 7 and below ("post-7"). Band 8 was undetectable in horse, cow and guinea pig, and was in higher amounts in rat. A band corresponding to a molecular weight of about 22 kD in the "post-8" region was present only in guinea pig RBC membranes.     

Preparation of neurotensin selectively iodinated on the tyrosine 3 residue. Biological activity and binding properties on mammalian neurotensin receptors

[Monoiodo- Tyr3 ]neurotensin, a neurotensin analogue that contains a single iodine atom on the side chain of Tyr 3 was prepared and purified. This analogue can be labeled at any specific radioactivity between 0 and 2000 Ci/mmol; its binding and biological properties on rat and guinea pig neurotensin receptors are identical to those of the parent peptide. These properties make [monoiodo- Tyr3 ]neurotensin the best suitable radioligand for detection and characterization of neurotensin receptors in various tissues and species.     

Somatostatin and VIP neurons in the retina of different species

Summary Neurons displaying somatostatin or vasoactive intestinal polypeptide (VIP) immunoreactivity were detected among the amacrine cells in the retina of baboon, cynomolgus monkey, squirrel monkey, cow, pig, cat, rabbit, guinea-pig, rat, mouse, frog and goldfish. Generally, immunoreactive cell bodies were located in the inner nuclear layer with processes ramifying in three more or less well-defined sublayers in the inner plexiform layer. The density of the sublayers and their location varied with the peptide and species investigated. In most cases there was a sublayer in the outermost part (Ramon y Cajal's sublamina 1) of the inner plexiform layer and this sublayer was usually the best developed. In some species a few somatostatin fibres were also detected in the outer plexiform layer, suggesting that some interplexiform cells contain somatostatin. In the baboon VIP was found exclusively in interstitial amacrine cells which have their cell bodies and processes entirely within the inner plexiform layer.