Expression of the homeobox engrailed gene during the embryonic development of the nervous system of the trout (Salmo fario L.).

The expression of the engrailed homeobox gene during trout embryogenesis has been examined using immunohistochemistry and the monoclonal antibody 'Mab 4D9'. Engrailed has been suggested to play an important role during development by controlling position-specific characteristics in the CNS of the early embryo. In the present study we have analyzed the expression of engrailed at 5 stages of embryonic development of the trout (Salmo fario L.). The earliest stage analyzed was when the optic vesicles appear. Engrailed was then expressed in the posterior mesencephalon and anterior metencephalon, in a caudorostrally decreasing gradient. As the embryo develops, the pattern of the engrailed expression increases in spatial complexity. Thus, in the later stages of development, just before hatching, engrailed was found in hypothalamic areas, the germinative matrix layer of the cerebellum, the mesencephalic tegmentum, the caudal optic tectum and in the area of the trigeminal motor nucleus. This is similar to the distribution of engrailed in embryos of amphibians, birds and mammals.     

Temporal disparity in pineal and retinal ontogeny

The development of photoreceptors and two putative neurotransmitter systems in the pineal organ and retina was studied during embryogenesis in the three-spined stickleback Gasterosteus aculeatus L. The investigation was performed by aid of immunocytochemistry using well characterized antisera to the retinal proteins alpha-transducin (TD alpha) and S-antigen (SA) (photoreceptor-markers), antisera against L-glutamic acid decarboxylase (GAD), gamma-aminobutyric acid (GABA), choline-O-acetyltransferase (ChAT) and with acetylcholinesterase (AChE) histochemistry (neurotransmitter-markers). It was possible to set up the following developmental time-table concerning the first appearance of positive immuno- and enzyme-reactive cells in the pineal organ and retina: I AChE-activity and TD alpha- and SA-immunoreactive cells in the pineal organ; II GAD- and GABA-immunoreactive cells in the pineal organ and retina; ChAT immunoreactivity and AChE activity in the retina; III hatching; IV SA-immunoreactive cells in the retina. The obtained results provide good evidence that while photoreceptor cells develop much earlier in the pineal organ than in the retina, neurons develop simultaneously in the pineal organ and retina.     

FMRFamide-like immunoreactive neurons of the nervus terminalis of teleosts innervate both retina and pineal organ

The tetrapeptide FMRFamide (Phe-Met-Arg-Phe-NH2) was first isolated from molluscan ganglia. Subsequently, it has become clear that vertebrate brains also contain endogenous FMRFamide-like substances. In teleosts, the neurons of the nervus terminalis contain an FMRFamide-like substance, and provide a direct innervation to the retina (Proc. Natl. Acad. Sci. U.S.A., 81 [1984] 940-944). Here we report the presence of FMRFamide-immunoreactive axonal bundles in the pineal organ of Coho salmon and three-spined sticklebacks. The largest numbers of axons were observed proximal to the brain, in the pineal stalk, while the distal part of the pineal organ contained only few axons. No FMRFamide-like-immunoreactive (IR) cell bodies were observed in the pineal organ. In adult fish it was not possible to determine the origin of these axons, due to the large numbers of FMRFamide-like IR axons in the teleost brain. However, by following the development of FMRFamide-like IR neurons in the embryonic and larval stickleback brain, it was possible to conclude that, at least in newly hatched fish, FMRFamide-like IR axons that originate in the nucleus nervus terminalis reach the pineal organ. Thus, it seems there is a direct connection between a specialized part of the chemosensory system and both the retina and the pineal organ in teleost fish.     

Detection of alterations in all three exons of the peripherin/RDS gene in Swedish patients with retinitis pigmentosa using an efficient DGGE system.

AIMS: To develop a sensitive mutation screening procedure suitable for routine analysis of the peripherin/RDS gene, and to estimate the nature and prevalence of peripherin/RDS gene mutations in Swedish patients with autosomal dominant retinitis pigmentosa. METHODS: To make the method as sensitive as possible, as many as eight segments, covering the three exons and the flanking intron sequences of the peripherin/RDS gene, were analysed by denaturing gradient gel electrophoresis. A group of 38 Swedish patients with a clinical diagnosis of autosomal dominant retinitis pigmentosa were screened for mutations in the peripherin/RDS gene. RESULTS: Three point mutations were found in four of the patients and five polymorphisms were defined. One mutation in exon 1, R172W, has been described previously in other ethnic groups as causing a macular degeneration. Another mutation, in exon 2 and causing the substitution F211L, was found in two unrelated patients. A third mutation, resulting in the likely non-pathogenic substitution S289L, as well as a polymorphism not reported previously, was found in exon 3. CONCLUSIONS: The screening procedure described allows detection of mutations in all of the exons, including the polymorphic 5' and 3' ends of the gene, and is therefore suitable for routine screening of peripherin/RDS gene defects in patients with autosomal dominant retinitis pigmentosa. The frequency of mutations found in the Swedish patient group indicates that defects in the peripherin/RDS gene might be a more common cause of autosomal dominant retinitis pigmentosa than was thought previously.     

Spermidine/spermine-N1-acetyltransferase activity in isoprenaline-stimulated rat salivary glands

In parotid, sublingual and submaxillary glands stimulated by continuous intravenous infusion of isoprenaline at various doses for 3 h, the concentrations of the polyamines putrescine, spermidine, spermine and N1-acetylspermidine as well as the activities of ornithine decarboxylase and spermidine/spermine-N1-acetyltransferase were determined. Ornithine decarboxylase is the enzyme that catalyses the formation of putrescine from ornithine. The most striking effect was observed in parotid glands. Here, the beta-adrenergic agonist increased the activity of spermidine/spermine-N1-acetyltransferase markedly, but only slightly (if at all) that of ornithine decarboxylase. It also increased the concentrations of N1-acetylspermidine and putrescine markedly. The result obtained in this gland indicates interconversion of higher polyamines to putrescine.     

Parasympathetic non-adrenergic, non-cholinergic-induced protein synthesis and mitogenic activity in rat parotid glands

Electrical stimulation of the parasympathetic auriculo-temporal nerve (40 Hz, 30 min), in the anaesthetized rat under - and -adrenoceptor blockade, increased [3H]thymidine and [3H]leucine uptake into the parotid glands by 80 and 263 %, respectively. The increase in response to parasympathetic stimulation was almost the same ([3H]thymidine 82 % and [3H]leucine 283 %) when atropine (2 mg kg-1 I.P. or I.V.) was included in the pretreatment. Neither intravenous infusion of vasoactive intestinal peptide (0.5-20 mg kg-1 min-1, 30 min) nor of bethanechol (10 mg kg-1 min-1, 30 min), under adrenoceptor blockade, increased the uptake of [3H]thymidine into the glands. However, these drugs increased [3H]leucine uptake, and in combination they interacted positively. Whereas vasoactive intestinal peptide is likely to be involved in the parasympathetic nerve-evoked protein synthesis, the nature of the non-adrenergic, non-cholinergic component(s) involved in the mitogenic response is presently unknown.     

Receptors involved in the nervous system regulation of polyamine metabolism in rat salivary glands

Polyamines are important for protein synthesis and tissue growth. In rat salivary glands, the activity of ornithine decarboxylase (ODC), the enzyme catalysing the formation of putrescine, and the content of putrescine, spermidine, spermine and N1-acetylspermidine were assayed after parasympathetic or sympathetic nerve stimulation in the presence of various autonomic receptor blockers. Increases in ODC activity occurred on activation of non-adrenergic and non-cholinergic receptors in response to parasympathetic nerve stimulation and on activation of alpha(alpha 1)- as well as of beta(beta 1)-adrenoceptors in response to sympathetic nerve stimulation. Moreover, in parotid glands, a beta(beta 1)-adrenoceptor-mediated inverse pathway for putrescine formation seemed to exist: from spermidine via N1-acetylspermidine.     

Antibody reactivity in penicillin-sensitive patients determinated with different penicillin derivatives.

35 individuals showing reactions to penicillin of anaphylactic shock, angioedema or urticaria were investigated. Their skin sensitivity was analysed using 16 different penicillin derivatives. In addition, the content of circulating reagins against the penicilloyl structure in the patient's sera were analysed using RAST. 17 of the patients had negative skin reactions and RAST results to all substances tested. The other 18 were skin test-positive to at least one derivative but showed markedly heterogeneous patterns of skin reactivity. 14 had positive reactions against penicilloyl structures accompanied by anti-penicilloyl reagins. Four patients showed doubtful reactions only to penicillin or penicilloate and/or penilloate. These patients also had very low levels of reagins against penicilloyl in their sera. Positive skin test results using monovalent penicillin derivatives such as penicillin, penicilloate, penilloate, penicilloyl amide, penicilloyl-formyl-lysine, penicillamine, which cannot form a multivalent antigen with penicillyol specificity, indicated formation of other derivatives of importance in penicillin allergy, e.g., penicillamine protein conjugates. Three patients showed skin reactions to ampicillin polymer and two to benzyl-penicillin polymer. The skin tests performed with the penicillin derivatives used do not seem to give more information on the sensitivity of the patients than does the RAST using penicilloyl structures.     

Effects of atropine or high frequency burst excitation on the composition of parasympathetic rat parotid saliva

In the presence of atropine continuous trains of impulses applied to the auriculo-temporal nerve at relatively high frequencies evoked a flow of parotid saliva that amounted to 5-10% of that before administration of the muscarinic blocker. The output of total protein, amylase, sodium and potassium decreased markedly. However, in terms of concentration protein and amylase increased, while sodium decreased and potassium was unchanged. When, in the absence of atropine, the continuous mode of stimulation was replaced by bursts of impulses of high frequencies the amount of saliva decreased. However, the concentration of protein, amylase and potassium increased, while the concentration of sodium decreased. The findings are discussed in relation to a possible peptidergic innervation of the secretory elements.