Growth zone of the juvenile goldfish retina revealed by fluorescent flat mounts

The retinas of juvenile goldfish (Carassius auratus) were processed for fluorescence microscope studies with flat mounts. The fish were reared in a laboratory aquarium for 1–3 months after either one class of retinal dopaminergic (DA) cells or indoleamine-accumulating (IA) cells had been destroyed by intravitreal injection of 6-hydroxydopamine (6-OHDA; 2.5 m?g) or 5,7-dihydroxytryptamine (5,7-DHT; 2.5 μg). The fish grew 10–12 cm in body length on the average for 3 months. The retinas were isolated at various survival times after pretreatment with 6-OHDA or 5,7-DHT. To improve detection of the cells, a mixture of noradrenaline (NA; 0.5 μg) and 5,6-DHT (0.5 μg) was intravitreally injected into the eyes 2 hr before enucleation. Retinal growth occurred by cell proliferation at the circumferential margin distal to the retinal center. In this process, DA and IA cells were added in concentric rings, with a rate of cell accretion of approximately 12 cells per day, estimated from the mean cell density in the growth zone (about 252 μg width) over a 3-month period. In order to compensate for loss of dopaminergic innervation induced by intravitreal 6-OHDA, the proliferation rate appeared to be facilitated significantly compared to that in normal retinas, and long and centrally extending processes were found to emerge from the DA cells formed earliest after intravitreal injection of 6-OHDA.     

Primary structures of multiple forms of cytochrome P-450 isozyme 2 derived from rabbit pulmonary and hepatic cDNAs

Rabbit pulmonary and hepatic mRNA was used to construct cDNA libraries that were screened with a cDNA probe (pf 3/46) to murine cytochrome P-450 homologous with rat cytochrome P-450b. Three types of cDNA clones were identified on the basis of restriction analysis with Bst EII. Two types of clones (B0 and B1) were present in a library constructed from pulmonary mRNA. The nucleotide sequence of B0 cDNA encodes a protein of 491 amino acids having a sequence identical to one reported for isozyme 2. The sequence derived from the b1 cDNA also contains 491 amino acids but differs from the B0 sequence at six positions. B1 clones were also obtained from a library constructed from hepatic mRNA isolated 12 hr after a single treatment with phenobarbital. A third type of clone (B2) was also obtained from this library, but no B0 clones were found. The sequence derived from the B2 cDNA contains 491 amino acids and differs from the B0 and B1 sequences at 11 and 15 positions, respectively. A second hepatic cDNA library, constructed from mRNA from the liver of a rabbit treated with phenobarbital daily for 4 days, contained B0 clones. The sequence of one of these was found to be identical to that of the pulmonary B0 clones.     

Changes in systemic circulation and organ blood flow during induced total spinal block

Total spinal block with lidocaine was performed in adult mongrel dogs, and circulatory changes or changes in organ blood flow were investigated. Dogs were divided into group A (5.1 mg.kg-1) and group B (10.0 mg.kg-1) according to the dose of lidocaine. During total spinal block, HR (79% and 78%), MAP (64% and 69%), LV dp/dt max (58% and 49%) and CI (67% and 67%) decreased significantly compared with the control values in groups A and B respectively. However, SVR was unchanged. The circulatory change occurred more rapidly in group B. Organ blood flow was measured in group A. Blood flow was measured in four areas during total spinal block. It was significantly decreased at liver and slightly decreased at cerebral cortex and medullary substance. However, renal cortex blood flow was not altered.     

Two autopsy cases of primary pituitary carcinoma

We studied two autopsy cases of primary pituitary carcinoma. Case-1. A 45 year old female was admitted on Oct. 4 1978, with a complaint of right homonymous hemianopsia. And diagnosis was pituitary adenoma. Partial removal of pituitary tumor was performed on Oct. 23 1978. She died on Dec. 5 1978 due to bleeding of gastrointestinal tract. Autopsy disclosed a pituitary carcinoma invading the left hypothalamus, mamillary body, optic and V cranial nerves, and mid brain as well as sphenoid bone. No extracranial metastasis was noted. Case-2. A 44 year old female with a history of acromegaly for 6 years was admitted with a complaint of headache on May 8 1976. She was diagnosed as having pituitary adenoma. The subtotal removal of pituitary tumor was performed on May 21 1976 and followed by 4500 rad irradiation. At this time, pathological diagnosis was eosinophilic adenoma. Seven years later, she complained of progressive right hearing disturbance, dysarthria and ataxic gait 1983. The second subtotal removal of pituitary tumor was performed with a diagnosis of recurrence of pituitary adenoma on Oct. 7 1983. After the operation, she complicated sepsis and died on Jan. 14 1984. An autopsy disclosed a pituitary carcinoma from residual pituitary gland, continuously extending to the subarachnoid space of the pons, and invading right cerebello-pontine angle and cerebellum. The histological examination revealed pituitary carcinoma with high pleomorphism and glioblastoma multiform-like feature were within the tumor.(ABSTRACT TRUNCATED AT 250 WORDS)     

5-hydroxytryptamine: its facilitative action on [3H]dopamine release from the retina

5-Hydroxytryptamine (5-HT) at 5 X 10(-4) M caused a two-fold increase in [3H]dopamine (DA) release from retinal particulate fractions of the carp (Cyprinus carpio). The 5-HT action was dose- and Ca2+-dependent. The three 5-HT agonists examined (5-methoxytryptamine, 5-methoxy-N,N-dimethyltryptamine and tryptamine) were more effective than 5-HT on [3H]DA release. 5,6-Dihydroxytryptamine was the strongest competitor among drugs tested for [3H ]DA uptake, but did not evoke any significant release of [3H]DA. Noradrenaline (or DA) at 5 X 10(-4) M produced a large increase in [3H]DA release from the particulate fractions, but its action was Ca2+-independent. The 5-HT-induced DA release could be seen in the frog retina but not in the rat retina, which does not contain indoleamine-accumulating cells. The results obtained strongly suggest that 5-HT stimulates [3H ]DA release through a receptor mechanism on DAergic terminals and modifies the DA-cell's function in the retina.     

Molecular characterization of the murine Coh locus: an amino acid difference at position 117 confers high and low coumarin 7-hydroxylase activity in P450coh.

Coumarin 7-hydroxylase (P450coh) and steroid 15 alpha-hydroxylase (P450(15 alpha) are encoded by members within the mouse 2A subfamily. Since P450coh activity is regulated by the Coh locus, we characterized P450coh cDNAs in strains having high coumarin 7-hydroxylase activity (CohH homozygote) including 129/J and DBA/2J, and compared them with P450coh cDNAs in low activity strains (CohL homozygote) C57BL/6J, C3H/HeJ and AKR/J. The nucleotide sequences of these two cDNAs differ by a single base, which results in an amino acid difference at position 117 (Val in P450cohH and Ala in P450cohL). The CohH phenotype exhibits approximately 10-fold greater Vmax and four-fold lower Km values than those in the CohL. Male 129 AKF1/J expresses approximately equal amounts of P450cohH and P450cohL mRNAs, associated with two Coh alleles. The levels of P450coh and P450(15 alpha) mRNAs in the F1 offspring suggested that a trans-acting factor(s) appeared to regulate the expressions of the P450 genes. A recent duplication in the ancestral mouse established the line of descent to P450(15 alpha) from the ancestral P450coh gene. During evolution, amino acid substitutions have selectively occurred at positions which alter the enzyme's substrate specificity and increase in the specific activity. Consistent with an important role of natural selection in the evolution of these genes is the relatively high nonsynonomous substitution rates on the P450(15 alpha) and the P450coh branches. As a result of these evolution events, the gene family consists of members which exhibit an extremely high degree of structural similarity, but very divergent hydroxylase activities and modes of regulation.